Skip to main content
PLOS One logoLink to PLOS One
. 2021 Mar 24;16(3):e0248949. doi: 10.1371/journal.pone.0248949

Prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical lesions and its associated factors among women aged 21–49 years in Amhara region, Northern Ethiopia

Minwuyelet Maru Temesgen 1,*, Tefera Alemu 1, Birtukan Shiferaw 1, Seid Legesse 1, Taye Zeru 2, Mahteme Haile 2, Tesfaye Gelanew 3
Editor: Ivan Sabol4
PMCID: PMC7990306  PMID: 33760866

Abstract

Background

Human papillomavirus (HPV) infection is considered as the major risk factor for the development of cervical cancer, second most frequent cancer in Ethiopia. However, the magnitude of the problem and the associated factors remain unrevealed in the Amhara region. This study aimed to determine the prevalence of HPV infection and factors contributing to the progression of HPV infection to cervical cancer.

Methods

Facility-based cross-sectional study design was employed among women aged 21 to 49 years of age who came for routine cervical cancer screening to 4 randomly selected hospitals (2 general and 2 referral) of Amhara region from May to October, 2019. The sample size was calculated by using the single population proportion formula, proportionated to hospitals, and women were recruited consecutively. Socio demographic and clinical data were collected using a pretested questionnaire and detection of HPV infection was done using HPV test (OncoE6TM Cervical Test) specific to HPV16/18 in cervical swabs. Visual inspection with acetic acid (VIA) was used to determine cervical lesions (precancerous and cancerous). Descriptive statistics and bivariate and multivariate logistic regression were used to describe HR-HPV and cervical lesions burden and association between HR-HPV, and cervical lesions and potential risk factors.

Results

Among 337 women 21 to 49 years (median age of 35 years ±SD = 7.1 years) of age enrolled in the study, The overall prevalence of oncogenic HPVs (HPV16/18) and the VIA-positivity rate, possible an indicative of cervical lesions, were 7.1% and 13.1%, respectively. Logistic regression analysis showed a significant association between early age of first sexual intercourse (COR = 2.26; 95% CI: 1.0–5.05) and level of education (COR = 0.31; 95% CI: 0.12–0.78) with cervical lesions. Higher odds of HPV positivity (COR = 1.56; 95% CI: 0.59–4.11, p = 0.36) and VIA positivity (COR = 1.39; 95% CI: 0.64–3.00, p = 0.39) were observed among participants who had a history of sexually transmitted illnesses (STIs).

Conclusions

There was a relatively low prevalence of oncogenic HPV 16/18 and VIA-positivity in women attending four hospitals in the Amhara Region. Early age sexual contact, high parity, and being uneducated/low level of education were independently associated factors with HR-HPV infection and development of cervical lesions, highlighting the importance of prioritizing the limited HPV testing to those risk groups.

Introduction

Human papillomavirus (HPV) comprises over 150 related DNA viruses that spreads sexually and through skin to skin contacts. Based on their virulence, these viruses are generally classified as high risk and low risk (LR) types [1]. Often our immunity can spontaneously clear off most of the HPV infections without treatment. However, infections with high risk HR-HPV types, especially in immune-compromised, can persists and possibility leads to warts, precancerous lesions or cancers [2]. That is why chronic infections with HR-HPV types are deep-rooted in almost all cervical cancer cases.

In the last six decades, the world has made significant progress towards the reduction of morbidity and mortality associated with cervical cancer. Despite this, cervical cancer remains a major threat to women’s health, especially in resource-limited countries due to poor awareness and late detection of cervical lesions. For instance, 85% of deaths from cervical cancer occurred in low and middle-income countries [3]. Besides, 14% of global incidences and 18% of deaths occurs in Sub-Saharan African countries [4].

In Ethiopia cervical cancer is a major cause of morbidity and mortality of women of age 15–49 years with prevalence ranging from 14% to 17% [57]. Several factors have been identified to be risk factors for developing cervical cancer associated with HPV infections. These factors can be but not limited to poor awareness, socioeconomic and, cultural factors, lack of primary and secondary prevention methods, poor nutrition and other sexual transmitted infections (STIs: HIV infection, trichomoniasis, syphilis, hepatitis B and hepatitis C).

The World Health Organization (WHO) 2013 cervical cancer guideline recommended a regular screening for woman of reproductive age using visual inspection with acetic acid (VIA) or when possible HPV testing followed by cryotherapy treatment, in resource- limited setting. VIA alone is widely used for presumptive diagnosis of cervical cancer in extreme resource-limited settings, where cervical cancer screening by cervical cytology and HPV testing is unavailable [8]. The Ethiopia cervical cancer prevention and control 2015 guideline indicated three components of cervical cancer prevention and control. The first is primary prevention which focuses on decreasing HPV infection through behavioral changes or vaccination. The secondary prevention targets on screening and treating precancerous lesions and tertiary care is the third which contains management of invasive cervical cancer [9]. Several studies have shown the likelihood of unvaccinated HPV-negative women to develop cervical cancer in the next 5–10 years is less, suggesting primary HPV testing is an essential preventive approach, particularly among unvaccinated women [10]. In Ethiopia, vaccination of girls prior to sexual debut using bivalent HPV vaccine that protects against HPV genotypes 16 and 18 was started among school girls as a pilot including Amhara region in 2019. However, the vaccines target certain genotypes and leave some proportion of the population infected by other genotypes unprotected. In addition, the vaccination coverage is very low and implementing this intervention is challenging in the regions where access to HPV-based testing is limited. Although, screening and treating cervical lesions using VIA remain crucial for successful cervical cancer prevention and control programs, it has some drawbacks related to sensitivity and specificity. Studies have shown VIA has about 67 to 79% sensitivity and 83 to 84% specificity [11, 12]. Therefore, screening with VIA together with HPV testing would be the best method for better outcome. Nonetheless, the proportion of HPV infection and cervical lesions (precancerous and cancerous lesions) among reproductive-age women as well as the associated factors remain unknown in the Amhara region despite this information is broadly seen as indispensable to the cervical cancer prevention programs. The main aim of this study was therefore to assess the magnitude of VIA positivity rate and prevalence of HR-HPV infection. The secondary aim was to identify associated potential risk factors to the development of precancerous or cancerous lesions.

Materials and methods

Study design and setting

A facility based cross-sectional study was conducted from May to October 2019 in four randomly selected government hospitals in the Amhara region. These hospitals were Tefera Hailu Memorial General Hospital, Felegehiwot Referral Hospital, Debretabor General Hospital and Debremarkos Referral Hospital that provides both primary HPV testing and cervical cancer screening and family planning services. Amhara Region is the second-largest populous region in Ethiopia which has 12 zones, 3 city administrations, and 180 districts (139 rural and 41 urban) and has 80 hospitals (5 referral, 2 general, and 73 primaries), 847 health centers, and 3,342 health posts. Screening women of reproductive age for cervical lesions using VIA in the region has been scaled up to all hospitals since 2015. However, HPV testing has been done in general and referral hospitals.

Study population

All women who came for routine gynecologic or family planning services to those hospitals were used as a source population for this study. Women of age 21 to 49 years who were referred to the routine cervical cancer screening services in the study period were invited to participate and consented after being screened by VIA and HPV testing. However, women with confirmed cancerous lesions and pregnancy were excluded from the present study to avoid unnecessary discomforts related to the procedure’s endocervical swabs.

Sample size and sampling technique

Sample size was determined using a single population proportion formula by considering 5% margin of error, 95% confidence level, 16% prevalence of HPV infection in Ethiopia, 5% non-response rate and 1.5 design effect and women were recruited consecutively [13]. The total sample size, 337, was proportionated to the selected participating hospitals based on their clients’ size: Tefera Hailu Memorial General Hospital (n = 51), Felegehiwot Referral Hospital (n = 118), Debretabor General Hospital (n = 65), and Debremarkos Referral Hospitals (n = 103).

Data collection

Ten data collectors (1 nurse and 1 laboratory technician) from each study site and four supervisors from Amhara Public Health Institute (APHI) were recruited. Scio-demographic and clinical data were collected by study nurses using a structured and pre-tested questionnaire through face to face interviewing each one of the participant women. The pre-test was conducted on 5% of the study population. All clients of reproductive age who were tested by routine VIA were invited to participate in this study, and were screened for HPV infection after being consented and interviewed.

Diagnosis of cervical lesions using visual inspection with acetic acid

After cleaning away any extra mucus with a cotton swab, a five percent acetic acid solution was applied to the cervix for VIA. After 30 to 60 seconds, abnormal cervical tissues (e.g., pre-cancerous or cancerous lesions) turned white [14]. On the other hand, no aceto white lesions was observed on the VIA of normal cervical tissue.

E6 HPV 16/18 oncoprotein testing

Endocervical swab was collected from study participant women by inserting swab devices into endocervix and rotating them counter-clockwise to make full circle three times. Detection of HPV types 16 and 18 in the cervical swabs was performed using E6 HPV 16/18 oncoprotein detection lateral flow (LF) strip test (OncoE6TM Cervical Test (Arbor Vita Corporation, Fremont, CA, USA) as per manufacturer’s instruction. Interpretation of the test result was also done according to the manufacturer’s instruction.

Data analysis

Collected data were checked for completeness before data entry. Data were cleaned, coded and entered in to Epidata 3.1 software and exported to STATA version 14 for analysis. Frequencies, proportion and summary statistics were used to describe the study population with relevant variables. Logistic regression was used to identify factors associated with HPV infection and odds ratio with 95% CI was used to assess the degree of association. P value <0.05 is considered statistically significant association and variables with P<0.2 were tested for multivariable logistic regression.

Ethical considerations

The study was approved by the Amhara Public Health Institute Ethical Review Board (Approval Number: 03/379/2011). Written permissions were obtained from zonal and district health authorities and the respective hospitals. All study participants were informed about the purpose of the study, and written consent (finger print for those who cannot read and write) was taken from each one of them. Confidentiality of any information related to the participants was maintained by excluding personal identifiers. Participants with a positive HPV test were linked to clinicians for further treatment and follow-up.

Results

Socio-demographic and clinical characteristics of participant women

A total of 337 women 21 to 49 years of age were enrolled in the study with median age of 35 years (±SD = 7.1 years). Of which 189 (56.1%) were rural residents and 182 (54.0%) participants were below primary education. Regarding occupational status 130 (38.6%) were house wives. The majority, 257 (76.3%) of participants were married whereas 45 (13.3%) were divorced (Table 1).

Table 1. Demographic characteristics of participant women (n = 337) aged 21–49 years, Amhara region, Ethiopia, 2019.

Characteristic Frequency (N = 337) Percent
Age in years
    21–30 112 33.2
    31–40 158 46.9
    41–49 67 19.9
Education
    Primary and below 182 54.0
    Secondary education 53 15.7
    College and above 102 30.3
Occupation
    House wife 130 38.6
    Employee 102 30.3
    Farmer 54 16.0
    Merchant 31 9.2
    Other 20 5.9
Marital status
    Never married 27 8.0
    Married 257 76.3
    Divorced 45 13.3
    Widowed 8 2.4
Residence
    Urban 148 43.9
    Rural 189 56.1

The mean number of children was 3.8 (±SD = 2.3) ranging from 1 to 9 with mean family size of 4.3 (±SD = 2.1) ranging from 1 to 13. All study participant women (n = 337) were screened for HIV and assessed for history of STIs when enrolled in this study and 67 (19.9%) and 61 (18.1%) of them had HIV and history of STIs, respectively. Two hundred fifty-seven (76.3%) of participants had a history of contraceptive use and of which 16.8% reported oral contraceptive use for 1 to 25 years with mean of 3.2 years (±SD = 3.9). Majority, 255 (75.7%) of study participants had heard about cervical cancer while 161 (47.8%) had not heard about symptoms of cervical cancer. Two hundred ninety-nine (88.7%) never heard about HPV and only 10 (2.9%) had got vaccinated for HPV. Most importantly, only 52 (15.4%) of study participants reported being screened for HPV infection (Table 2).

Table 2. Awareness of cervical cancer among participant women (n = 337) aged 21–49 years, Amhara region, Ethiopia, 2019.

Characteristic Frequency (N = 337) Percent
Heard about cervical cancer
    Yes 255 75.7
    No 82 24.3
Heard about symptoms of cervical cancer
    Yes 176 52.2
    No 161 47.8
Heard about HPV
    Yes 299 88.7
    No 38 11.3
Vaccinated for HPV
    Yes 10 2.9
    No 327 97.1
Screened for HPV infection
    Yes 52 15.4
    No 285 84.6

HR-HPV infection, cervical lesions and associated factors

All the 337 participant women had successful HPV and VIA tests. Of which 24 (7.12%) and 44 (13.05%) women had positivity to HR-HPV and VIA tests, respectively. As shown in Tables 3 and 4, the proportion of HPV infection and VIA-positivity was shown to be evenly distributed across different age groups, however, relatively higher HPV and VIA positivity rates were observed among older participants (41–49 years).

Table 3. Proportion of HPV infection and cervical lesions among participant women (n = 337) aged 21–49 years, Amhara region, Ethiopia, 2019.

Characteristic Number (N = 337) HPV infection based on E6 16/18 antigen test VIA cervical cancer screening
Positive Negative Positive Negative
N (%) N (%) N (%) N (%)
Age in years
    21–30 112 8 (7.1) 104 (92.9) 9 (8.1) 103 (91.9)
    31–40 158 9 (5.7) 149 (94.3) 18 (11.4) 140 (88.6)
    41–49 67 7 (10.4) 60 (89.6) 17 (25.4) 50 (74.6)
Overall 337 24 (7.1) 313 (92.9) 44 (13.1) 293 (86.9)
Education
    Primary and below 182 15 (8.2) 167 (91.8) 30 (16.5) 152 (83.5)
    Secondary education 53 7 (13.2) 46 (86.8) 8 (15.1) 45 (84.9)
    College and above 102 2 (1.9) 100 (98.1) 6 (5.9) 96 (94.1)
Overall 337 24(7.1) 313(92.9) 44(13.1) 293(86.9)
Marital status
    Never married 27 5 (18.5) 22 (81.5) 4 (14.8) 23 (85.2)
    Married 257 14 (5.5) 243 (94.5) 27 (10.5) 230 (89.5)
    Divorced 45 3 (6.7) 42 (93.3) 10 (22.2) 35 (77.8)
    Widowed 8 2 (25.0) 6 (75.0) 3 (37.5) 5 (62.5)
Residence
    Urban 148 11 (7.4) 137 (92.6) 16 (10.8) 132 (89.2)
    Rural 189 13 (6.9) 176 (93.1) 28 (14.8) 161 (85.2)

Table 4. Prevalence of HPV (16/18) infections among participant women (n = 337) aged 21–49 years, with and without cervical lesions, Amhara region, Ethiopia, 2019.

Tissue type Number (N = 337) HPV positive HPV negative
Cervical lesions 44 17 (38.6%) 27 (61.4%)
No cervical lesions 293 7 (2.4%) 286 (97.6%)

The prevalence of HR-HPV 16/18 infections among women with and without cervical lesions was 38.6% and 2.4%, respectively (Table 4). Moreover, VIA identified 17 and missed 7 out of 24 HPV positive women that make its sensitivity 70.8%. Although its positive predictive value is 61.4% (27 out of 44), 286 out of 293 participant women were correctly identified negative which make VIA sensitivity 91.3% The prevalence of HR-HPV 16/18 infections in HIV-negative and HIV-positive women was 8.1% and 3.0%, respectively (Table 5).

Table 5. Logistic regression analysis of HPV positivity among participant women (n = 337) aged 21–49 years, Amhara region, Ethiopia, 2019.

Characteristic Number (N = 337) HPV infection based on E6 16/18 antigen test COR (95% CI) p-value AOR (95%CI) p-value
Positive n (%) Negative n (%)
Age
    21–30 112 8 (7.1) 104 (92.9) 1
    31–40 158 9 (5.7) 149 (94.3) 0.78 (0.29–2.10) 0.63
    41–49 67 7 (10.4) 60 (89.6) 1.51 (0.52–4.39) 0.44
Age of first intercourse
    ≥18 106 4 (3.8) 102 (96.2) 1
    <18 231 20 (8.7) 211(91.3) 2.41 (0.80–7.25) 0.11 1.31 (0.37–4.61) 0.66
HIV
    Negative 270 22 (8.1) 248 (91.9) 1
    Positive 67 2 (3.0) 65 (97.0) 0.34 (0.07–1.51) 0.15 0.31 (0.06–4.46) 0.14
STI history
    No 276 18 (6.5) 258 (93.5) 1
    Yes 61 6 (9.8) 55 (90.2) 1.56 (0.59–4.11) 0.36

Multivariate logistic regression analysis revealed slightly higher odds of HPV infection in study participants who had history of STIs, and who started first sexual contact when they were under 18 years of age as compared to their counter parts (Table 5).

Significant association between cervical lesions with educational status and age of first sexual intercourse was observed using multivariate logistic regression analysis. Participant women with college degree or higher education had 0.31 reduced odds of cervical lesions than women who had primary education or less. Women who had started sexual contact at age of less than 18 years old had 2.26 times more likely to have cervical lesions than their counter parts (Table 6). The level of education was also found to be significantly associated with knowledge of cervical cancer indicated by chi square (p<0.001). One hundred fourteen (62.6%) women having primary education and below had heard about cervical cancer as compared to those women who had secondary school (84.9%) and college (94.12%).

Table 6. Logistic regression analysis of VIA positivity among participant women (n = 337) aged 21–49 years, Amhara region, Ethiopia, 2019.

Characteristic Number (N = 337) VIA positivity COR (95% CI) p-value AOR (95%CI) p-value
Positive n (%) Negative n (%)
Residence
    Urban 148 16 (10.8) 132 (80.2) 1
    Rural 189 28 (14.8) 161 (85.2) 1.43 (0.74–2.76) 0.28
Education
    Primary & below 182 30 (16.5) 152 (83.5) 1 1
    Secondary 53 8 (15.1) 45 (84.9) 0.90 (0.38–2.10) 0.80 1.43 (0.11–18.17) 0.78
    College & above 102 6 (5.9) 96 (94.1) 0.31 (0.12–0.78) 0.01 1.13 (0.01–110.24) 0.95
Age of first intercourse
    ≥18 106 8 (7.6) 98 (92.4) 1
    <18 231 36 (15.6) 195 (84.4) 2.26 (1.0–5.05) 0.04 1.88 (0.05–64.95) 0.72
Parity
    0 35 5 (14.3) 30 (85.7) 1
    1–2 110 10 (9.1) 100 (90.9) 0.6 (0.19–1.89) 0.38
    3–4 92 8 (8.7) 84 (91.3) 0.57 (0.17–1.88) 0.35
    ≥5 100 21(21.0) 79 (79.0) 1.59 (0.55–4.61) 0.38
HIV status
    Negative 270 32 (11.9) 238 (88.1) 1 1
    Positive 67 12 (17.9) 55 (82.1) 1.62 (0.78–3.35) 0.19 5.47 (0.26–111.49) 0.26
STI history
    No 276 34 (12.3) 242 (87.7) 1
    Yes 61 10 (13.4) 51 (83.6) 1.39 (0.64–3.00) 0.39

Logistic regression analysis also revealed higher odds of cervical lesions among study participants who had HIV-positive status 17.9% vs 11.8%; COR = 1.62; 95% CI: 0.78–3.35) and had history of STIs (13.4% vs 12.3%; COR = 1.39; 95% CI: 0.64–3.00). In addition, slightly higher proportion of cervical lesions was also observed among study participants who resided in rural areas (18.8% vs 10.8%), and had ≥5 children (parity) (21% vs 8.7% of 3–4, 9.10% of 1–2 and 14.21 of null parity).

Discussion

In this study we assessed HPV 16/18 infections, the presence of cervical lesions and associated factors among women aged 21 to 49 years in four hospitals in the Amhara region, Ethiopia. The overall prevalence of HPV and cervical lesions (precancerous and cancer lesions) in the study area was 7.1% and 13.1%, respectively. The observed HPV prevalence in the present study is lower compared to studies in another part of Ethiopia, by Leyh-Bannurah S. et al., 17.3% [15], and by Ruland R. et al. in Attat hospital of rural Ethiopia, 16% [7]. This could mainly due to considerable differences in HPV detection methods between us and others. The antigen detection method we used detects only HPV 16 and 18 types, whereas others used molecular detection method for several HPV types. Also, the prevalence of presumed cervical lesion is lower than studies reported by Gedefaw A. et al. from southern Ethiopia 16.5% [16]. This is possibly due to the fact we excluded pregnant women and women with cervical cancers from our study. In support of this, studies show that HPV is the most common cause of viral cervical infection in pregnancy [1720].

Globally, HPV types 16 and 18 predominate and are responsible for most anogenital HPV-related cancers in women [21]. In our finding HPV prevalence among women without cervical lesions was 2.4% which is comparable with studies in Sudan (3.2%) [22], but much lower than other East Africa countries, Mozambique (40.3%) [23] and Kenya (41.4%) [24].

HPV prevalence among participant women with cervical lesions was 38.6%. Our finding is lower than from previous reports by Bekele A. et al. and Gemechu A. et al. in Ethiopia: Jimma (67.1%) and Hawasa (49.3%) [25, 26]. The lower prevalence of HPV in the present study can be justified as women of cervical cancer complaint visiting family planning or gynecology clinics were excluded from the present study. However, the proportion of VIA positivity in the present study is 13.1%, which is consistent with findings from different parts of Ethiopia, Addis Ababa, 13.1%, [27], Debremarkos, 14.1% [6] and Hawassa,12.9% [28].

The sensitivity and specificity of VIA in our study was 70.8% and 91.3% respectively which is low but comparable with studies by Arbyn M. et al. and Bobdey S. et al. [11, 12]. However, in our finding 7 out of 24 VIA negative women were identified positive by HPV OncoE6TM test which might indicate the importance of HPV testing together with VIA screening for early detection and follow up.

In accordance with the study by Abate S. et al. in Addis Ababa, Ethiopia [29], the prevalence of HPV infection and cervical lesions in the current study was similar across the different age groups with a peak in the age range of 40 to 49 years. These studies including ours suggest age is not an independent associated factor for HPV persistence infection, however, older women are more likely to develop persistent HR-HPV infections and cervical lesions [30].

Susceptibility to HPV infection is dependent on exposure but also on characteristics of the host such as immunosuppression and the presence of HIV. The present study, however, unlike studies from Zambia [31] and Rwanda [32] revealed the low prevalence of HPV (3%) among HIV positive women than women without HIV (8%). This finding could be explained partly by exclusion of women with cervical cancer from the study. Also, the most common HR- HPV types among HIV-positive women would be non-HPV 16/18 types that could not be detected by the commercial HPV test kit used in the present study. On the contrary, HIV-positive participants had 1.62 times higher odds to develop cervical lesions than HIV negative, supporting HIV is an independent factor for cervical cancer [33]. Without antiretroviral therapy, HIV infection often leads to chronic immunosuppression, which in turn results in persistent HPV infection, a prerequisite to progression to cervical lesions. Nevertheless, given the small number of HIV-positive participants in the present study, future study on HIV-positive women is required to provide intervention strategies that would much benefit the HIV-positive women.

Unsafe sex is a primary transmission of genital human HPV, and STIs is an indicative of having unsafe sex. Concurrent to this, we found a slightly higher prevalence of HPV infections among women who had history of STIs (9.8%) than who had not (6.5%). The possible explanations might be because women co-infected with non-HPV STIs like chlamydia trachomatis, herpes simplex or genital warts could have altered epithelial tissue due to local inflammatory response which in turn make those women more susceptible to persistent HPV infection [34, 35].

Other cofactors are known to contribute for progression from cervical HPV infection to cervical cancer. In the present study, low level of education or being uneducated and high parity were found as associated factor for development of cervical lesions. Additionally, we found an association of cervical lesions with rural residence and history of STIs, albeit the association was not as such strong. In support of this, high proportion of uneducated and rural participants in this study never heard about cervical cancer, suggesting presence of health seeking behavior and knowledge gap in the region. A similar finding was reported from different studies in Ethiopia [3638]. Women who had history of STIs were found 1.39 times higher odds to develop cervical lesions than women had no history of STIs. Similar finds were reported in studies from other regions of the Ethiopia [39].

Studies from Ethiopia, Africa and other parts of the world show that first sexual contact before age of 18 years is risk factor for cervical cancer [4042]. Surprisingly, high proportion (69%) of participants in the present study reported having first sexual contact before the age 18 years, and of which 16% were diagnosed to have cervical lesions by VIA screening. This suggest early age sexual contact is also associated factor for cervical cancer, and the need to vaccinate female children at age 7–9 years before they have first sexual contact in an attempt to significantly reduce women deaths and mortality related to cervical cancer.

Conclusions

Our study shows that prevalence of oncogenic HPV 16/18 and cervical lesions among women visiting hospitals for gynecology and family planning services are low. This is possibly because of (i) exclusion of pregnant women with confirmed cervical cancer and pregnancy from the study, and (ii) poor sensitivity of VIA. The present study indicates testing for HPV together with VIA improves early detection of high risk women for successful cervical cancer screening programs. However, more research is needed to better understand VIA positivity among HPV infected women. This study also identifies early age sexual contact related to early marriage, high parity and low level of education as independent associated factors to develop cervical lesions, supporting the importance of prioritizing the limited HPV screening test to those risk groups as well as vaccinating female children at age of 7–9 years.

Limitation of the study

There are limitations in this study. First, the prevalence of cervical lesions in our study is more likely underestimated for the fact that high risk women were excluded from study, and possibly due to the low sensitivity of VIA to identify cervical lesions. Second, it is possible that our results regarding HPV infections are underestimated as the HPV E6 16/18 antigen detection test, employed in this study, detects only HPV 16 and 18 types and the low number of women enrolled in the study.

Supporting information

S1 Dataset. Dataset for prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical lesions and its associated factors among women aged 21–49 years in Amhara region, Northern Ethiopia.

(XLSX)

Acknowledgments

We would like to thank study participants, staffs of study hospitals and Amhara Public Health Institute (APHI) staffs who had been providing constructive comments and technical support.

List of abbreviations

APHI

Amhara Public Health Institute

HIV

Human Immunodeficiency Virus

HPV

Human Papillomavirus

HR

High risk

LR

Low risk

STI

Sexually transmitted illness

VIA

Visual Inspection with Acetic acid

WHO

World Health Organization

Data Availability

All relevant data are within the manuscript and its Supporting Information files.

Funding Statement

The authors received no specific funding for this work.

References

  • 1.Walboomers J., et al., Human papilloma virus is a necessary cause of invasive cervical cancer worldwide. J Pathol 1999. 189: p. 12–19. [DOI] [PubMed] [Google Scholar]
  • 2.WHO. Human papilloma virus and cervical cancer. 2016. [cited 2019 January, 15]. [Google Scholar]
  • 3.Denny L. and Prendiville W., Cancer of the cervix: Early detection and cost-effective solutions. Int J Gynaecol Obstet, 2015. 131 Suppl 1: p. S28–32. 10.1016/j.ijgo.2015.02.009 [DOI] [PubMed] [Google Scholar]
  • 4.De Vuyst H., et al., The burden of human papillomavirus infections and related diseases in sub-saharan Africa. Vaccine, 2013. 31 Suppl 5: p. F32–46. 10.1016/j.vaccine.2012.07.092 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Hailemariam T., et al., Prevalence of Cervical Cancer and Associated Risk Factors among Women Attending Cervical Cancer Screening and Diagnosis Center at Yirgalem General Hospital, Southern Ethiopia. J Cancer Sci Ther 2017. 9(11). [Google Scholar]
  • 6.Getinet M., et al., Prevalence and predictors of Pap smear cervical epithelial cell abnormality among HIV-positive and negative women attending gynecological examination in cervical cancer screening center at Debre Markos referral hospital, East Gojjam, Northwest Ethiopia. BMC Clin Pathol, 2015. 15: p. 16. 10.1186/s12907-015-0016-2 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Ruland R., et al., Prevalence of human papilloma virus infection in women in rural Ethiopia. Eur J Epidemiol, 2006. 21(9): p. 727–9. 10.1007/s10654-006-9055-4 [DOI] [PubMed] [Google Scholar]
  • 8.WHO, Prevention of cervical cancer through screening using visual inspection with acetic acid (VIA) and treatment with cryotherapy. 2012. [Google Scholar]
  • 9.Health F.M.o. Guideline for Cervical Cancer Prevention and Control in Ethiopia. 2015. [cited 2020; Available from: https://www.iccp-portal.org/system/files/plans/Guideline%20Eth%20Final.pdf. [Google Scholar]
  • 10.Denny L. and Anorlu R., Cervical cancer in Africa. Cancer Epidemiol Biomarkers Prev, 2012. 21(9): p. 1434–8. 10.1158/1055-9965.EPI-12-0334 [DOI] [PubMed] [Google Scholar]
  • 11.Arbyn M., et al., Pooled analysis of the accuracy of five cervical cancer screening tests assessed in eleven studies in Africa and India. International journal of cancer, 2008. 123(1): p. 153–160. 10.1002/ijc.23489 [DOI] [PubMed] [Google Scholar]
  • 12.Bobdey S., et al., Burden of cervical cancer and role of screening in India. Indian journal of medical and paediatric oncology: official journal of Indian Society of Medical & Paediatric Oncology, 2016. 37(4): p. 278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Gebremariam T., Human papillomavirus related cervical cancer and anticipated vaccination challenges in Ethiopia. International Journal of Health Sciences, Qassim University, 2016. 10(1). [PMC free article] [PubMed] [Google Scholar]
  • 14.Ardahan M. and Temel A.B., Visual inspection with acetic acid in cervical cancer screening. Cancer Nursing, 2011. 34(2): p. 158–163. 10.1097/NCC.0b013e3181efe69f [DOI] [PubMed] [Google Scholar]
  • 15.Leyh-Bannurah S.-R., et al., Cervical human papillomavirus prevalence and genotype distribution among hybrid capture 2 positive women 15 to 64 years of age in the Gurage zone, rural Ethiopia. Infectious agents and cancer, 2014. 9: p. 33. 10.1186/1750-9378-9-33 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Gedefaw A., Astatkie A., and Tessema G.A., The prevalence of precancerous cervical cancer lesion among HIV-infected women in southern Ethiopia: a cross-sectional study. PLoS One, 2013. 8(12): p. e84519. 10.1371/journal.pone.0084519 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Castellsagué X., et al., Human Papillomavirus (HPV) infection in pregnant women and mother-to-child transmission of genital HPV genotypes: a prospective study in Spain. BMC infectious diseases, 2009. 9(1): p. 74. 10.1186/1471-2334-9-74 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Chang-Claude J., et al., Longitudinal study of the effects of pregnancy and other factors on detection of HPV. Gynecologic Oncology, 1996. 60(3): p. 355–362. 10.1006/gyno.1996.0055 [DOI] [PubMed] [Google Scholar]
  • 19.Dongrui D., et al., Asymptomatic genital infection of human papillomavirus in pregnant women and the vertical transmission route. Journal of Huazhong University of Science and Technology [Medical Sciences], 2005. 25(3): p. 343–345. [DOI] [PubMed] [Google Scholar]
  • 20.Dinc B., et al., Prevalence of human papillomavirus (HPV) and HPV-16 genotyping by real-time PCR in patients with several cervical pathologies. Brazilian Journal of Infectious Diseases, 2010. 14(1): p. 19–23. [DOI] [PubMed] [Google Scholar]
  • 21.Chan C.K., et al., Human Papillomavirus Infection and Cervical Cancer: Epidemiology, Screening, and Vaccination—Review of Current Perspectives. Journal of Oncology, 2019. 2019: p. 3257939. 10.1155/2019/3257939 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Salih M.M., et al., Genotypes of human papilloma virus in Sudanese women with cervical pathology. Infect Agent Cancer, 2010. 5: p. 26. 10.1186/1750-9378-5-26 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Naucler P, et al., Human papillomavirus type-specific risk of cervical cancer in a population with high human immunodeficiency virus prevalence: case-control study. J Gen Virol. 2011. 92(12): p. 2791. [DOI] [PubMed] [Google Scholar]
  • 24.De Vuyst H, et al., The prevalence of human papillomavirus infection in Mombasa, Kenya. Cancer Causes Control, 2010. 21: p. 2313. 10.1007/s10552-010-9645-z [DOI] [PubMed] [Google Scholar]
  • 25.Bekele A., et al., Human papillomavirus type distribution among women with cervical pathology—a study over 4 years at Jimma Hospital, southwest Ethiopia. Trop Med Int Health, 2010. 15(8): p. 890–3. 10.1111/j.1365-3156.2010.02552.x [DOI] [PubMed] [Google Scholar]
  • 26.Gemechu Ameya and Yerakly F., Characteristics of cervical disease among symptomatic women with histopathological sample at Hawassa University referral hospital, Southern Ethiopia. BMC Women’s Health, 2017. 17(91). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Kirubel Eshetu Ali, et al., Burden and genotype distribution of highrisk Human Papillomavirus infection and cervical cytology abnormalities at selected obstetrics and gynecology clinics of Addis Ababa, Ethiopia. BMC Cancer, 2019. 19(768). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Zewdie Mulissa Deksissa, Fessahaye Alemseged Tesfamichael, and Henok Assefa Ferede, Prevalence and factors associated with VIA positive result among clients screened at Family Guidance Association of Ethiopia, south west area office, Jimma model clinic, Jimma, Ethiopia 2013: a cross-sectional study. BMC Res Notes 2015. 8(618). 10.1186/s13104-015-1594-x [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.Abate S.M., Trends of Cervical Cancer in Ethiopia. Cervical Cancer 2016. 1(1). [Google Scholar]
  • 30.Kang L.-N., et al., A prospective study of age trends of high-risk human papillomavirus infection in rural China. BMC infectious diseases, 2014. 14(1): p. 96. 10.1186/1471-2334-14-96 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Sahasrabuddhe V.V., et al., Prevalence and distribution of HPV genotypes among HIV-infected women in Zambia. Br J Cancer, 2007. 96(9): p. 1480–3. 10.1038/sj.bjc.6603737 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Mukanyangezi M., et al., Persistence rate of cervical human papillomavirus infections and abnormal cytology in Rwanda. HIV medicine, 2019. 20(7): p. 485–495. 10.1111/hiv.12782 [DOI] [PubMed] [Google Scholar]
  • 33.Branca M., et al., Assessment of risk factors and human papillomavirus (HPV) related pathogenetic mechanisms of CIN in HIV-positive and HIV-negative women. Study design and baseline data of the HPV-PathogenISS study. European journal of gynaecological oncology, 2004. 25(6): p. 689–698. [PubMed] [Google Scholar]
  • 34.Silva J., Cerqueira F., and Medeiros R., Chlamydia trachomatis infection: implications for HPV status and cervical cancer. Archives of gynecology and obstetrics, 2014. 289(4): p. 715–723. 10.1007/s00404-013-3122-3 [DOI] [PubMed] [Google Scholar]
  • 35.Ghosh I., et al., Association of genital infections other than human papillomavirus with pre-invasive and invasive cervical neoplasia. Journal of clinical and diagnostic research: JCDR, 2016. 10(2): p. XE01. 10.7860/JCDR/2016/15305.7173 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Bayu H., et al., Cervical Cancer Screening Service Uptake and Associated Factors among Age Eligible Women in Mekelle Zone, Northern Ethiopia, 2015: A Community Based Study Using Health Belief Model. PLoS One, 2016. 11(3): p. e0149908. 10.1371/journal.pone.0149908 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Aweke Y.H., Ayanto S.Y., and Ersado T.L., Knowledge, attitude and practice for cervical cancer prevention and control among women of childbearing age in Hossana Town, Hadiya zone, Southern Ethiopia: Community-based cross-sectional study. PLOS ONE, 2017. 12(7): p. e0181415. 10.1371/journal.pone.0181415 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Ali K.E., et al., Burden and genotype distribution of high-risk Human Papillomavirus infection and cervical cytology abnormalities at selected obstetrics and gynecology clinics of Addis Ababa, Ethiopia. BMC cancer, 2019. 19(1): p. 768. 10.1186/s12885-019-5953-1 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Wolday D., et al., HPV genotype distribution among women with normal and abnormal cervical cytology presenting in a tertiary gynecology referral Clinic in Ethiopia. Infectious agents and cancer, 2018. 13(1): p. 28. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Panjaliya R., Dogra V., and Gupta S., Study of the risk factors associated with cervical cancer. Biomedical and Pharmacology Journal, 2015. 3(1): p. 179–182. [Google Scholar]
  • 41.Adewuyi S., Shittu S., and Rafindadi A., Sociodemographic and clinicopathologic characterization of cervical cancers in northern Nigeria. European journal of gynaecological oncology, 2008. 29(1): p. 61. [PubMed] [Google Scholar]
  • 42.Kassa R.T., Risk factors associated with precancerous cervical lesion among women screened at Marie Stops Ethiopia, Adama town, Ethiopia 2017: a case control study. BMC research notes, 2018. 11(1): p. 1–5. 10.1186/s13104-017-3088-5 [DOI] [PMC free article] [PubMed] [Google Scholar]

Decision Letter 0

Ivan Sabol

9 Nov 2020

PONE-D-20-26948

Prevalence of oncogenic human papilloma virus (HPV 16/18) infection, cervical cancer and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia

PLOS ONE

Dear Dr. Temesgen,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

==============================

According to the expert reviews provided the manuscript would require additional revisions before being suitable for publication.

Please address the comments of individual reviewers with special emphasis on study participant selection, about the questionnaire administered, about the ethical questions and the description of the statistical analysis since all are a big part of the PLOS Publication criteria

https://journals.plos.org/plosone/s/criteria-for-publication

Furthermore, some additional issues to be addressed are listed below

==============================

Please submit your revised manuscript by Dec 24 2020 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

  • A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.

  • A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.

  • An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

We look forward to receiving your revised manuscript.

Kind regards,

Ivan Sabol

Academic Editor

PLOS ONE

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. We suggest you thoroughly copyedit your manuscript for language usage, spelling, and grammar. If you do not know anyone who can help you do this, you may wish to consider employing a professional scientific editing service.  

Whilst you may use any professional scientific editing service of your choice, PLOS has partnered with both American Journal Experts (AJE) and Editage to provide discounted services to PLOS authors. Both organizations have experience helping authors meet PLOS guidelines and can provide language editing, translation, manuscript formatting, and figure formatting to ensure your manuscript meets our submission guidelines. To take advantage of our partnership with AJE, visit the AJE website (http://learn.aje.com/plos/) for a 15% discount off AJE services. To take advantage of our partnership with Editage, visit the Editage website (www.editage.com) and enter referral code PLOSEDIT for a 15% discount off Editage services.  If the PLOS editorial team finds any language issues in text that either AJE or Editage has edited, the service provider will re-edit the text for free.

Upon resubmission, please provide the following:

  • The name of the colleague or the details of the professional service that edited your manuscript

  • A copy of your manuscript showing your changes by either highlighting them or using track changes (uploaded as a *supporting information* file)

  • A clean copy of the edited manuscript (uploaded as the new *manuscript* file)

3. Thank you for including your ethics statement:

'The study was approved by APHI ethical review board ref. number 03/379/2011. All study subjects during the study period were informed about the purpose of the study and their consent was sought in written. Confidentiality of any information related to the participants was maintained using code numbers. Participants with a positive HPV test were linked to clinicians for further treatment and follow up.'

a. Please amend your current ethics statement to include the full name of the ethics committee/institutional review board(s) that approved your specific study.

b. Once you have amended this/these statement(s) in the Methods section of the manuscript, please add the same text to the “Ethics Statement” field of the submission form (via “Edit Submission”).

For additional information about PLOS ONE ethical requirements for human subjects research, please refer to http://journals.plos.org/plosone/s/submission-guidelines#loc-human-subjects-research

Additional Editor Comments:

1) It seems each VIA positive patient is considered as having cervical cancer. This doesn't appear to be accurate since VIA would be positive for pre-cancer lesions as well which the authors acknowledge at line 80. Please consider rephrasing the relevant text to include pre-cancer to VIA results for example at page 10 Line 168 and elsewhere

2) Line 128 which variables were included in the multivariable logistic regression?

3) Table 2 „Vaccinated for cc“ should be „Vaccinated for HPV“

4) Line 170 typo „HVP“ should be HPV

5) Line 174 and elsewhere please rephrase „abnormal cervical cytology“ to „cervical lesions“ or something similar since cytology was not performed

6) Tables 5 & 6 why were different variables considered for HPV or VIA positivity?

7) Table 6 the numbers for „Residence“ do not add up to 337

8) Sentence at line 223-224 is unclear. Possibly the authors meant „cervical cancer complaint“?

9) Language and grammar should be somewhat improved

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Partly

**********

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

**********

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

**********

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: Yes

**********

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Reviewer’s comments

Full title: Prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical cancer and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia

Manuscript number: PONE-D-20-26948

Corresponding author: Minwuyelet Maru Temesgen Amhara Public Health Institute Dessie Branch Dessie, Amhara Region ETHIOPIA

Comments and questions

Overall comments

I would like to thank the authors for addressing potential factors for the emergence of health issues; cervical cancer among reproductive-age women in developing nations. Therefore, evidence regarding the prevalence of HPV is important to inform decision-making policy dealing with the prevention and control program of cervical cancers. The study presents the findings of original research in the area of women’s health and the results have not been published elsewhere. The study didn’t adequately perform relevant analyses with sufficient detail. The discussion part must comprehensively be written addressing policy, practical, methodological, and scientific implications. The article is presented not an acceptable level of English language standard and reporting guidelines.

Additional comments and questions

The author would respond to some of the following questions and comments.

1. Title: “Prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical cancer and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia”. The “…oncogenic human papillomavirus (HPV 16/18) infection…” is one of the risk factors for cervical cancer. But it's independently or separately mentioned in the topic. In the topic; there also “… it's associated factors…” though this is not clear to which condition these factors are considered to be associated. So, the author would restructure the title as…; “Prevalence of cervical cancer and its associated factors…”.

2. Abstract: Methods: Line 25: They would discuss sample calculation, sampling techniques, method of analysis used to address the prevalence and associated factors.

3. Abstract: conclusion: line 41 to 43: “…This study also identifies early age sexual contact, high parity and being uneducated/low educational status as primary risk factors to the development of cervical cancer…”. The study design of this study was “cross-sectional”, but the author tried to conclude the finding as “risk factors”. The risk factor is not a good expression or not appropriate for such a study design. Therefore; the author would replace phrases like “…associated factors”. See the topic.

4. Introduction: page: Line 75-78: “…In the Amhara region, testing for HPV infection has been not available or if available it is in limited areas, and hence little is known about the prevalence of HPV infection and cervical cancer, their co-prevalence’s and factors contributing to HPV infection and progression to cervical cancer”. The author mentioned the lack of HPV tests in the region as a justification to conduct the current study. This is not a good reason to conduct this study because the study has nothing to do with the availability or lack of HPV test facilities. Surprisingly; the study is conducted where the HPV test facilities are available. Can the author respond to this question?

5. Methods and materials: Study setting: Line 81: the author would briefly discuss the information the study settings that address health care system/health services in the region (health coverage, No.hospital, No. health center, coverage of cervical cancer care centers, HPV tests facilities, etc.)

6. Methods and materials: Study setting: Line 86-90: “All women who came for routine gynecologic or family planning services to those hospitals were used as a source population for this study. Women 88 age 21 to 49 years and referred to the cervical cancer screening services during the study period were included in the study. However, women with known cervical cancer and pregnancy were excluded”. This statement is talking about the study population and misplaced here. Therefore; the author would move it to the “study population section”.

7. Methods and materials: Sample size and sampling technique: Line 91: the authors should clarify how they draw/select the participants.

8. Methods and materials: data collection: Line 98: the interview technique must be clarified. Was it at the entry point or exit point?

9. Result part: How did the authors handle the relationship between HPV and cervical cancer?

10. Result: Table 5 (line 191), table 6 (line 201): “bivariate logistic regression…” what does it mean by “bivariate logistic regression”?. Since you are doing with multiple factors affecting the outcome variable?

11. Discussion: Line 213: “…The observed HPV prevalence in the present study is lower compared to studies in another part of Ethiopia, Gurage zone, 17.3%[12], and Atta hospital of rural Ethiopia, 16% [7]. Also, the prevalence of presumed cervical cancer is lower than studies reported from southern Ethiopia 16.5% [13]”. The authors tried to justify reasons as “…Knowledge about the prevalence of HPV among women with normal and abnormal cervical cytology is important to monitor and design HPV control programs…”. What does it mean? Did you explored the knowledge of women in this regard? If “yes”, was it high or low? Comment: the author would discuss the implication for health policy and practices. E.g the lower prevalence in your finding may be due to the inaccessibility of the services.

Reviewer #2: In general, I found this study interesting and relevant to the field. It is well designed and follow an appropriate methodology and analysis. However, there is unclarity on the consent procedure and participant selection to accept this manuscript as it is. I hope, the suggestions and questions below will help the authors to improve their manuscript.

Comments

Remove ‘this’ line 79

Q1, Please, clearly explain the inclusion and exclusion criteria. Line 87-90, “women age 21-49 years referred to the cervical cancer screening services during the study…”

Was this cervical screening service for the purpose of study or it was given as a routine service at all the four included hospitals?

Did the participant send it to the cervical cancer screening test after consent or before?

where was consent taken? Was at ANC, PMTCT, FP, or at the cancer screening center?

Q2, Ethical considerations, and consent procedures should be included in the body of the manuscript.

Q3, Participants also screened for HIV. What does screening mean? Were the participants tested for HIV? Or asked for self-reporting their HIV status? If tested, did they counseled? who performed the testing? How did you manage refusals?

Q4, why did you use a design effect in this study? The design effect is used for multistage sampling studies. But this study is a facility-based cross-sectional study.

Q5, Modify table name for table 1. – “Demographic characteristics of study participants, women aged 21-49 years…”,. The Demographic characteristics are not about the whole Amahar region. Follow an appropriate table naming and apply this for all other tables.

Q6, Remove the total raw end of table 1. It is confusing and it is better to put n=337 at the headers of the table as Frequency(N= 337). Apply this for all other tables.

Q7, Line 151, replace “had not heard” by never heard about

Q8, Line 224: “visiting family planning or gynecology clinics 224 were excluded from the present study” Was this included in the exclusion criteria? And justify the reason why the women were excluded.

**********

6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Fira Abamecha

Reviewer #2: Yes: Serebe Gebrie

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

PLoS One. 2021 Mar 24;16(3):e0248949. doi: 10.1371/journal.pone.0248949.r002

Author response to Decision Letter 0


13 Dec 2020

Response to the Editor and Reviewers:

Dear Editor and Reviewers!

The authors would like to thank the area editor and the reviewers for their precious time and invaluable comments to our submitted article “Prevalence of oncogenic human papilloma virus (HPV 16/18) infection, cervical cancer and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia”. We greatly appreciate the thorough and thoughtful comments provided to our paper. We are resubmitting the revised version according to the comments and we believe that the revised version of our paper addresses all concerns. Minor revisions and grammatical corrections are applied to the manuscript modification along with revisions explained in this document. Attached below are detailed responses to the comments.

Response to the Editor

1) It seems each VIA positive patient is considered as having cervical cancer. This doesn't appear to be accurate since VIA would be positive for pre-cancer lesions as well which the authors acknowledge at line 80. Please consider rephrasing the relevant text to include pre-cancer to VIA results for example at page 10 Line 168 and elsewhere

Response: It is addressed

• VIA can be positive in precancerous or cancerous lesions and therefore pre-cancer lesion is added to relevant section in the revised version.

2) Line 128 which variables were included in the multivariable logistic regression?

Response: We appreciate the query. Variables with p-value less than 0.2 in binary logistic regression were included in multivariable logistic regression. Based on this we have now mentioned as indicated in line 138 of the revised manuscript

• For association of HPV acquisition; age of first sexual intercourse, HIV status, occupation and marital status were included in multivariable logistic regression

• For association of VIA positivity; age, age of first sexual intercourse, HIV status, educational status, occupation, time of oral contraceptive use and marital status were included in multivariable logistic regression

3) Table 2 „Vaccinated for cc“ should be „Vaccinated for HPV“

Response: Corrected

4) Line 170 typo „HVP“ should be HPV

Response: Corrected

5) Line 174 and elsewhere please rephrase „abnormal cervical cytology“ to „cervical lesions“ or something similar since cytology was not performed

Response: Thank you for this important comment. We rephrase abnormal cervical cytology to cervical lesions

6) Tables 5 & 6 why were different variables considered for HPV or VIA positivity?

Response: The variables are selected based on the assumption and certain evidences that factors for acquiring HPV infection are different from factors for developing cervical cancer. Therefore, variables in table 5 (current age, age of first sexual intercourse, HIV status and history of sexually transmitted illness) are related to sexual behavior which could affect acquisition of HPV, whereas, variables in table 6 (age of first sexual intercourse, HIV status, residence, history of STI, educational status and parity) were considered as factors for cervical cancer.

7) Table 6 the numbers for „Residence“ do not add up to 337

Response: It is addressed

8) Sentence at line 223-224 is unclear. Possibly the authors meant „cervical cancer complaint“?

Response: It is addressed

9) Language and grammar should be somewhat improved

• It is done

Response to Reviewer #1

1. Title: “Prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical cancer and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia”. The “…oncogenic human papillomavirus (HPV 16/18) infection…” is one of the risk factors for cervical cancer. But it's independently or separately mentioned in the topic. In the topic; there also “… it's associated factors…” though this is not clear to which condition these factors are considered to be associated. So, the author would restructure the title as…; “Prevalence of cervical cancer and its associated factors…”.

Response: Thank you for this suggestion. As the reviewer points out, oncogenic human papilloma virus infection is one of the risk factor for cervical cancer; however our study was also aimed to describe the prevalence of HPV with associated factors for infection. By considering this, we believe the title as it is would be more explanatory.

2. Abstract: Methods: Line 25: They would discuss sample calculation, sampling techniques, method of analysis used to address the prevalence and associated factors.

Response: Thank you for this comment. It is addressed

• The sample calculation used was single population proportion formula and women were consecutively recruited (page 2, line 29, line111). Sixteen percent HPV prevalence, 5% margin of error and 1.5 design effect was considered for the calculation. (Line 111)

3. Abstract: conclusion: line 41 to 43: “…This study also identifies early age sexual contact, high parity and being uneducated/low educational status as primary risk factors to the development of cervical cancer…”. The study design of this study was “cross-sectional”, but the author tried to conclude the finding as “risk factors”. The risk factor is not a good expression or not appropriate for such a study design. Therefore; the author would replace phrases like “…associated factors”. See the topic.

Response: It is addressed.

• Risk factor is replaced with associated factor

4. Introduction: page: Line 75-78: “…In the Amhara region, testing for HPV infection has been not available or if available it is in limited areas, and hence little is known about the prevalence of HPV infection and cervical cancer, their co-prevalence’s and factors contributing to HPV infection and progression to cervical cancer”. The author mentioned the lack of HPV tests in the region as a justification to conduct the current study. This is not a good reason to conduct this study because the study has nothing to do with the availability or lack of HPV test facilities. Surprisingly; the study is conducted where the HPV test facilities are available. Can the author respond to this question?

Response: We sincerely appreciate the comment. The reviewer mention lack of HPV test or availability in limited area cannot be justification to conduct the current study.

• We would like to mean due in many health facilities of the region, the prevalence remain unknown (line 84) and also indicated in the method. The current study provided HPV test kit for the study purpose to selected hospitals which have HPV testing setups (testing machine). Although the study has nothing to do (or plan to do) with the availability of HPV tests in health facilities, by revealing the facts about the prevalence and associated factors, it recommend to scale up HPV testing to every facility in the region.

5. Methods and materials: Study setting: Line 81: the author would briefly discuss the information the study settings that address health care system/health services in the region (health coverage, No.hospital, No. health center, coverage of cervical cancer care centers, HPV tests facilities, etc.)

Response: We acknowledge this information pointed out by the reviewer

• We added description of the study setting, the health care system and cervical cancer services in the region to the study setting ( Line 95)

6. Methods and materials: Study setting: Line 86-90: “All women who came for routine gynecologic or family planning services to those hospitals were used as a source population for this study. Women 88 age 21 to 49 years and referred to the cervical cancer screening services during the study period were included in the study. However, women with known cervical cancer and pregnancy were excluded”. This statement is talking about the study population and misplaced here. Therefore; the author would move it to the “study population section”.

Response: It is addressed and moved to study population (Line 101

7. Methods and materials: Sample size and sampling technique: Line 91: the authors should clarify how they draw/select the participants.

Response: It is addressed.

• The sample size is calculated as indicated in comment 2 and study participants were recruited consecutively

8. Methods and materials: data collection: Line 98: the interview technique must be clarified. Was it at the entry point or exit point?

Response: We apologize for our lack of clarity. It is clarified in the revised version (Line 115).

• All eligible clients were screened for cervical lesions using VIA and tested for HPV. Study participants were recruited at the exit point for interview and review of medical records to get VIA and HPV results after getting informed consent.

9. Result part: How did the authors handle the relationship between HPV and cervical cancer?

Response: It is an established fact that HPV infection is a risk factor for cervical cancer. In our study we focused on describing the prevalence of oncogenic HPV and cervical cancer or precancerous lesions separately with possible associated factors for HPV acquisition and cervical lesions, thus HPV and cervical cancer association was not described.

10. Result: Table 5 (line 191), table 6 (line 201): “bivariate logistic regression…” what does it mean by “bivariate logistic regression”?. Since you are doing with multiple factors affecting the outcome variable?

Response: We thank the reviewer for pointing out the type of analysis in relation to factors affecting the outcome variable. We have considered the comment and both crude odds ratio (COR) and adjusted odds ratio (AOR) was used to describe the association in table 5 (Line 215) and table 6 (Line 246)

11. Discussion: Line 213: “…The observed HPV prevalence in the present study is lower compared to studies in another part of Ethiopia, Gurage zone, 17.3%[12], and Atta hospital of rural Ethiopia, 16% [7]. Also, the prevalence of presumed cervical cancer is lower than studies reported from southern Ethiopia 16.5% [13]”. The authors tried to justify reasons as “…Knowledge about the prevalence of HPV among women with normal and abnormal cervical cytology is important to monitor and design HPV control programs…”. What does it mean? Did you explored the knowledge of women in this regard? If “yes”, was it high or low? Comment: the author would discuss the implication for health policy and practices. E.g the lower prevalence in your finding may be due to the inaccessibility of the services.

Response: The sentence “…Knowledge about the prevalence of HPV….” was intended for description of knowledge about HPV among women with and without cervical lesion and it is another concept not put to justify the low prevalence of HPV, thus was deleted for its ambiguity and the reason for low prevalence is justified; variation in detection methods ( in line 260), exclusion of pregnant women and cervical cancer cases (264).

Response to Reviewer #2

Comments

Remove ‘this’ line 79

Response:- It is done

Q1, Please, clearly explain the inclusion and exclusion criteria. Line 87-90, “women age 21-49 years referred to the cervical cancer screening services during the study…”

Was this cervical screening service for the purpose of study or it was given as a routine service at all the four included hospitals?

Did the participant send it to the cervical cancer screening test after consent or before?

where was consent taken? Was at ANC, PMTCT, FP, or at the cancer screening center?

Response: Thank you for pointing this out.

• Cervical screening service was given as a routine service in all hospitals. But for the purpose of the study HPV test kit was provided and all eligible clients screened for cervical lesions were also sent to test for HPV. Study participants were recruited at the exit point, after getting the service (VIA and HPV test). Consent was sought for interview and review of medical records to get VIA and HPV and HIV results. (Data collection: Line 115).

• The inclusion criteria: Women of age 21 to 49 years eligible for cervical cancer screening visiting the clinics for routine services during the study period were included (Line 104)

• Exclusion criteria: women with known cervical cancer and pregnancy were excluded (line 106).

Q2, Ethical considerations, and consent procedures should be included in the body of the manuscript.

Response: It is done (Line 333)

Q3, Participants also screened for HIV. What does screening mean? Were the participants tested for HIV? Or asked for self-reporting their HIV status? If tested, did they counseled? who performed the testing? How did you manage refusals?

Response: HIV testing is part of cervical cancer screening program and the result is obtained from medical records. The ethical issue was addressed in the consent.

Q4, why did you use a design effect in this study? The design effect is used for multistage sampling studies. But this study is a facility-based cross-sectional study.

Response: Thank you for this comment. Since the hospitals are found in different zonal administrations of the region, they are considered as clusters. Therefore, even though the study was cross sectional, design effect was used.

Q5, Modify table name for table 1. – “Demographic characteristics of study participants, women aged 21-49 years…”,. The Demographic characteristics are not about the whole Amahar region. Follow an appropriate table naming and apply this for all other tables.

Response: It is done

Q6, Remove the total raw end of table 1. It is confusing and it is better to put n=337 at the headers of the table as Frequency(N= 337). Apply this for all other tables.

Response: it is done

Q7, Line 151, replace “had not heard” by never heard about

Response: It is done

Q8, Line 224: “visiting family planning or gynecology clinics 224 were excluded from the present study” Was this included in the exclusion criteria? And justify the reason why the women were excluded.

Response: We apologize for our lack of clarity. The exclude ones were women of cervical compliant visiting the clinics. It is corrected

Attachment

Submitted filename: Response to Reviewers.docx

Decision Letter 1

Ivan Sabol

9 Feb 2021

PONE-D-20-26948R1

Prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical lesions and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia

PLOS ONE

Dear Dr. Temesgen,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

While most of the original comments were addressed, some additional issues were noted that should be improved or at least the limitations highlighted.

Please submit your revised manuscript by Mar 26 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

  • A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.

  • A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.

  • An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

We look forward to receiving your revised manuscript.

Kind regards,

Ivan Sabol

Academic Editor

PLOS ONE

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

Reviewer #4: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Partly

Reviewer #4: Partly

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

Reviewer #3: I Don't Know

Reviewer #4: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: No

Reviewer #3: Yes

Reviewer #4: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: Revised manuscript review

The authors have addressed all my concerns point-by-point and made a considerable revision. The manuscript is now greatly improved, most of the suggestions are included, and other issues are satisfactorily justified. However, minor editorial corrections are still needed but can be considered for publication after proofreading and minor editorial correction have done without further review of the revised version.

For example,

Abstract page 2, L29-31, there is a repetition of the whole sentence.

L94 – avoid spacing before a comma,

L73-74 – Revise the sentence ending with "in all most all cervical cancer cases".

L351-449 – Check reference writing(the font and spacing) as per the journal requirement. Unlike the body of the paper, the reference seems written in smaller font and without spacing.

Reviewer #3: Major concern concerning this study

In the Introduction section, the authors should elaborate in what consist the cervical cancer prevention programs in Ethiopia and provide relevant references. What is the status of cervical screening, VIA, HPV testing, and HPV vaccination? What is the particularity of Amhara region of Northern Ethiopia compared to the rest of the country? This study did not bring any particular knowledge that is already established in many other countries. So, the authors should focus on a particular goal and emphasize what novelty this study brings to the scientific community. In addition, the major limitation of the study is the low number of enrolled women (N=337), therefore the conclusion is unreadable. The authors should enlarge they study to at least few thousands respondents.

Minor, but essential, correction to do

Abstract, Methods, line 23-27. Unnecessary repetition.

Introduction, line 64: “and late detection of HPV infection” should be “and late detection of cervical lesion”

Introduction, line 86: What does it stand for “cervical cancer prevention programs”?

Introduction, line 79: “where HPV testing is unavailable” should be “where cervical cancer screening by cervical cytology and HPV testing is unavailable”

Results: all numbers should be with one decimal.

Discussion: when referring to others studies be precise, i.e. Author et al. When comparing your method of testing to others be precise as well and cite the appropriate reference.

Reviewer #4: The focus of the study article is very important to Ethiopia and other developing countries still using VIA as a cervical cancer screening strategy. However, the uptake of VIA is low and there are a lot of issues associated with its sensitivity and specificity. Besides, quality control is another grey area in the implementation of VIA as a primary screening strategy. Onco6 HPV antigen test is one of the HPV tests available in the market with high specificity for lesions associated with HPV 16 and 18 but its sensitivity is low as a test and this is the inherent problem of the test. As close to 70% of cervical cancer is associated with HPV16 and 18 the test is assumed to be one of the good options in the provision of HPV based cervical cancer screening service. However, as the purpose of cervical cancer screening is to identify women with pre-cancerous lesion and viruses other than HPV 16 and 18 maybe important in the study settings thus this particular study will miss significant number of high-risk HPV infections. Thus, with the test type employed the authors will be able to identify only persistent HPV16 and 18 infection and not the prevalence of high-risk HPV and this needs to be corrected in their text. Although VIA is advocated for its specificity by those who recommend the test for primary screening, VIA missed seven out of 24 Onco6 test positives and 27 of VIA positives were Onco6 test negatives. Of course, about seven of the 27 Onco6 test negatives had STI history. So many factors account for the progression of the HPV infection to cervical cancer, HPV genotype is one and they were only able to identify HPV 16 and 18 and not others by the method they have employed. As cervical cancer is a kind of progressive disease identifying factors associated with progression of HPV infection cannot be met by such correctional study design. In summary, the finding of this study is publishable if the authors rather show how VIA is neither a sensitive nor a specific test to identify pre-cancerous lesions. The authors need further to look at their data justify why 61% (27/44) of the VIA positive were Onco6 negative and recommend further study to assess the utility of VIA to identify specific pre-cancerous lesions among those who are Onco6 test positives.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: Yes: Serebe Gebrie

Reviewer #3: No

Reviewer #4: Yes: Tamrat Abebe

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

PLoS One. 2021 Mar 24;16(3):e0248949. doi: 10.1371/journal.pone.0248949.r004

Author response to Decision Letter 1


4 Mar 2021

Response to the Reviewers:

Dear Reviewers!

The authors would like to thank the reviewers for their precious time and vital comments to our submitted article titled “Prevalence of oncogenic human papilloma virus (HPV 16/18) infection, cervical lesions and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia” (PONE-D-20-26948R1). We greatly appreciate the comments and we are resubmitting the revised version. We believe the revised version addresses all concerns and attached below are detailed responses to the comments.

Response to Reviewer #2

1. Abstract page 2, Line 29-31, there is a repetition of the whole sentence

Response: - sorry for the repetition, It is corrected

2. Line 94: Avoid spacing before comma

Response: Thank you for this comment. It is addressed

3. Line 73-74 Revise the sentence ending with “in all most all cervical cancer cases”.

Response: It is addressed and corrected as “in almost all cervical cancer cases”

4. Line 351-499: Reference writing (font and spacing)

Response:- Thank you, the font and spacing is corrected

Response to Reviewer #3

• Cervical cancer prevention programs in Ethiopia and the status of VIA, HPV testing and vaccination in Ethiopia and Amhara region should be elaborated. In addition, low number of enrolled women is major limitation

Response: Thank you for this point.

The content of cervical cancer prevention and control program in Ethiopia is included in the introduction based on 2015 Ethiopian Ministry of health cervical cancer prevention and control guideline in Line 80, Line 87-96 of the revised manuscript and the low sample size is written in the limitation of the study.

• Abstract Method line 23-27, unnecessary repetition

Response: It is addressed

• Introduction line 64 “and late detection of HPV infection” should be “and late detection of cervical lesion”

Response: It is corrected

• Introduction line 86 what does it stand for “cervical cancer prevention program”

Response: Thank you for this comment. It is elaborated based on the local context.

• Introduction line 79: “where HPV testing is unavailable” should be “where cervical cancer screening by cervical cytology and HPV testing is unavailable”

Response: It is done

• Result: all numbers should be in one decimal

Response: it is done

• Discussion: when referring to others studies be precise, Author et al. When comparing your method of testing to others be precise as well and cite the appropriate reference

Response: It is addressed

Response to Reviewer #4

The focus of the study article is very important to Ethiopia and other developing countries still using VIA as a cervical cancer screening strategy. However, the uptake of VIA is low and there are a lot of issues associated with its sensitivity and specificity. Besides, quality control is another grey area in the implementation of VIA as a primary screening strategy. Onco6 HPV antigen test is one of the HPV tests available in the market with high specificity for lesions associated with HPV 16 and 18 but its sensitivity is low as a test and this is the inherent problem of the test. As close to 70% of cervical cancer is associated with HPV16 and 18 the test is assumed to be one of the good options in the provision of HPV based cervical cancer screening service. However, as the purpose of cervical cancer screening is to identify women with pre-cancerous lesion and viruses other than HPV 16 and 18 maybe important in the study settings thus this particular study will miss significant number of high-risk HPV infections. Thus, with the test type employed the authors will be able to identify only persistent HPV16 and 18 infection and not the prevalence of high-risk HPV and this needs to be corrected in their text. Although VIA is advocated for its specificity by those who recommend the test for primary screening, VIA missed seven out of 24 Onco6 test positives and 27 of VIA positives were Onco6 test negatives. Of course, about seven of the 27 Onco6 test negatives had STI history. So many factors account for the progression of the HPV infection to cervical cancer, HPV genotype is one and they were only able to identify HPV 16 and 18 and not others by the method they have employed. As cervical cancer is a kind of progressive disease identifying factors associated with progression of HPV infection cannot be met by such correctional study design. In summary, the finding of this study is publishable if the authors rather show how VIA is neither a sensitive nor a specific test to identify pre-cancerous lesions. The authors need further to look at their data justify why 61% (27/44) of the VIA positive were Onco6 negative and recommend further study to assess the utility of VIA to identify specific pre-cancerous lesions among those who are Onco6 test positives.

Response: Thank you for pointing out issues related to

- persistent HPV16 and 18 infection versus high-risk HPV

- the uptake, sensitivity and quality control of VIA,

- the importance of HPV genotypes other than 16 and 18,

- persistent HPV 16 and HPV 18 infection versus prevalence of high-risk HPV

The sensitivity and specificity of VIA was calculated and added to the result section (line 213-216). VIA identified 17 and missed 7 out of 24 HPV positive women that make its sensitivity 70.8% and 286 out of 293 participant women were correctly identified negative which make VIA specificity 91.3%. However, the positive predictive value of VIA is only 61.4% (27 out of 44). The low sensitivity of VIA is indicated in the limitation and further study is recommended (Line 348)

Regarding the HPV genotypes our study aimed to HPV 16 and HPV 18 which are considered highly prevalent genotypes associated with cervical cancer. Since women visiting gynecology and family planning service were recruited for the study, it is believed to be prevalence.

Attachment

Submitted filename: Response to Reviewers.docx

Decision Letter 2

Ivan Sabol

9 Mar 2021

Prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical lesions and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia

PONE-D-20-26948R2

Dear Dr. Temesgen,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Ivan Sabol

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

During revision some typos were introduced or missed. Please address these in particular and/or any other language or typographical issues.

Line 84 „prior to sexual debut using was started“ - missing words

Line 142 „OncoE6TM“ TM should probably be in superscript throughout the manuscript. For the test the manufacturer "Arbor Vita Corporation, Fremont, CA, USA" should be listed, not a commercial vendor.

Line 93 "Nonthless," typo

Line 143 „manufacture’s“ typo

Line 237 "who were educated above college had" - grammatically incorrect replace with "with college degree or higher education had"

Line 238 "educated primary and below." - grammatically incorrect replace with "women who had primary education or less".

Line 268 "13.4% vs12.3%;" no space between vs and the number

Line 299 „onco6 test“ is should be stated as before as „OncoE6 test“

Reviewers' comments:

Acceptance letter

Ivan Sabol

11 Mar 2021

PONE-D-20-26948R2

Prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical lesions and its associated factors among women aged 21-49 years in Amhara region, Northern Ethiopia.

Dear Dr. Temesgen:

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org.

If we can help with anything else, please email us at plosone@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Ivan Sabol

Academic Editor

PLOS ONE

Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    S1 Dataset. Dataset for prevalence of oncogenic human papillomavirus (HPV 16/18) infection, cervical lesions and its associated factors among women aged 21–49 years in Amhara region, Northern Ethiopia.

    (XLSX)

    Attachment

    Submitted filename: Response to Reviewers.docx

    Attachment

    Submitted filename: Response to Reviewers.docx

    Data Availability Statement

    All relevant data are within the manuscript and its Supporting Information files.


    Articles from PLoS ONE are provided here courtesy of PLOS

    RESOURCES