Abstract
Many diagnostic methods for sexually transmitted infections (STIs) have been developed. Because various infection agents are associated with STIs, and because infected persons sometimes show no symptoms, the diagnosis of STIs using nucleic acid amplification tests(NAATs) has required not only simultaneous multi-targeting, but also sensitive detection. Here, we compare microarray and real-time PCR for the detection of three common STIs agents, Ureaplasma urealyticum, Mycoplasma genitalium, and Chlamydia trachomatis, using human urine samples. The detection results showed that microarray and real-time PCR technology are both effective tools for the detection of STI agents. In conclusion, real-time PCR detection offers more sensitivity and specificity than microarray, because of the quantitative method employed. But, microarray offers better performance, in terms of high-throughput and simultaneous multi-targeting.
Keywords: STIs, Real-time PCR, Microarray, DNA chip, Molecular diagnostics
References
- 1.Takahashi S, et al. Detection of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum DNAs in urine from asymptomatic healthy young Japanese men. J. Infect. Chemother. 2006;12:269–271. doi: 10.1007/s10156-006-0462-y. [DOI] [PubMed] [Google Scholar]
- 2.Suntoke TR, et al. Evaluation of multiplex real-time PCR for detection of Haemophilus ducreyi, Treponema pallidum, herpes simplex virus type 1 and 2 in the diagnosis of genital ulcer disease in the Rakai District, Uganda. Sex. Transm. Infect. 2009;85:97–101. doi: 10.1136/sti.2008.034207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3.Safaeian M, et al. Comparison of the SPF10-LiPA system to the Hybrid Capture 2 Assay for detection of carcinogenic human papillomavirus genotypes among 5,683 young women in Guanacaste, Costa Rica. J. Clin. Microbiol. 2007;45:1447–1454. doi: 10.1128/JCM.02580-06. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Menton JF, et al. Molecular epidemiology of sexually transmitted human papillomavirus in a self referred group of women in Ireland. Virol. J. 2009;6:112. doi: 10.1186/1743-422X-6-112. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.Jensen JS, et al. Use of TaqMan 5′ nuclease real-time PCR for quantitative detection of Mycoplasma genitalium DNA in males with and without urethritis who were attendees at a sexually transmitted disease clinic. J. Clin. Microbiol. 2004;42:683–692. doi: 10.1128/JCM.42.2.683-692.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6.Shi G, et al. Fabrication and optimization of the multiplex PCR-based oligonucleotide microarray for detection of Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum. J. Microbiol. Methods. 2005;62:245–256. doi: 10.1016/j.mimet.2005.02.017. [DOI] [PubMed] [Google Scholar]
- 7.Cimino GD, et al. More false-positive problems. Nature. 1990;345:773–774. doi: 10.1038/345773b0. [DOI] [PubMed] [Google Scholar]
- 8.De Leeneer K, et al. Rapid and sensitive detection of BRCA1/2 mutations in a diagnostic setting: comparison of two high-resolution melting platforms. Clin. Chem. 2008;54:982–989. doi: 10.1373/clinchem.2007.098764. [DOI] [PubMed] [Google Scholar]
- 9.Kim CJ, et al. HPV oligonucleotide microarray-based detection of HPV genotypes in cervical neoplastic lesions. Gynecol. Oncol. 2003;89:210–217. doi: 10.1016/S0090-8258(02)00069-0. [DOI] [PubMed] [Google Scholar]
- 10.Weinstock H, Berman S, Cates W., Jr. Sexually transmitted diseases among American youth: incidence and prevalence estimates, 2000. Perspect. Sex. Reprod. Health. 2004;36:6–10. doi: 10.1363/3600604. [DOI] [PubMed] [Google Scholar]
- 11.Swygard H, et al. Targeted screening for Trichomonas vaginalis with culture using a two-step method in women presenting for STD evaluation. Sex. Transm. Dis. 2004;31:659–664. doi: 10.1097/01.OLQ.0000143091.95094.73. [DOI] [PubMed] [Google Scholar]
- 12.Almeda J, et al. Evaluation of a commercial enzyme immunoassay for HIV screening in urine. Eur. J. Clin. Microbiol. Infect. Dis. 2004;23:831–835. doi: 10.1007/s10096-004-1221-6. [DOI] [PubMed] [Google Scholar]
- 13.Rogers SM, et al. NAAT-identified and self-reported gonorrhea and chlamydial infections: different atrisk population subgroups? Sex. Transm. Dis. 2002;29:588–596. doi: 10.1097/00007435-200210000-00005. [DOI] [PubMed] [Google Scholar]
- 14.Kong F, et al. Comparative analysis and serovar-specific identification of multiple-banded antigen genes of Ureaplasma urealyticum biovar 1. J. Clin. Microbiol. 1999;37:538–543. doi: 10.1128/jcm.37.3.538-543.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Kong F, et al. Phylogenetic analysis of Ureaplasma urealyticum-support for the establishment of a new species, Ureaplasma parvum. Int. J. Syst. Bacteriol. 1999;49(Pt4):1879–1889. doi: 10.1099/00207713-49-4-1879. [DOI] [PubMed] [Google Scholar]
- 16.Jensen JS, Borre MB, Dohn B. Detection of Mycoplasma genitalium by PCR amplification of the 16S rRNA gene. J. Clin. Microbiol. 2003;41:261–266. doi: 10.1128/JCM.41.1.261-266.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17.Yoo HS, et al. DNA barcoding Korean birds. Mol. Cells. 2006;22:323–327. [PubMed] [Google Scholar]
- 18.Thorne LB, et al. Analytic validation of a quantitative real-time PCR assay to measure CMV viral load in whole blood. Diagn. Mol. Pathol. 2007;16:73–80. doi: 10.1097/PDM.0b013e318033ab9e. [DOI] [PubMed] [Google Scholar]
- 19.Morandi L, et al. Monitoring HCV RNA viral load by locked nucleic acid molecular beacons real time PCR. J. Virol. Methods. 2007;140:148–154. doi: 10.1016/j.jviromet.2006.11.014. [DOI] [PubMed] [Google Scholar]
- 20.Keyaerts E, et al. Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR. Int. J. Infect. Dis. 2006;10:32–37. doi: 10.1016/j.ijid.2005.02.003. [DOI] [PMC free article] [PubMed] [Google Scholar]