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. 1990 Dec;28(12):2739–2743. doi: 10.1128/jcm.28.12.2739-2743.1990

Rapid detection of human papillomavirus in cervical scrapes by combined general primer-mediated and type-specific polymerase chain reaction.

A J van den Brule 1, C J Meijer 1, V Bakels 1, P Kenemans 1, J M Walboomers 1
PMCID: PMC268265  PMID: 2177751

Abstract

A two-step polymerase chain reaction (PCR) procedure was used as a new screening strategy for the detection of human papillomavirus (HPV) genotypes in cervical scrapes omitting prior DNA extraction. Sample preparation consisted of a freeze-thaw step followed by boiling the cells before the PCR mixture was added. This pretreatment was as efficient and reproducible for HPV DNA amplification as DNA purification. By using crude cell suspensions, a prescreening of the samples with the general primer-mediated PCR method (GP-PCR) was performed to detect a broad spectrum of sequenced and still unsequenced HPV types at the subpicogram level. HPV-containing scrapes by GP-PCR were subjected to HPV 6, 11, 16, 18, 31, and 33 type-specific PCR (TS-PCR) to identify the sequenced HPV types. This direct GP/TS-PCR method was tested on a large group of cervical scrapes (n = 459) from women visiting a gynecologic outpatient clinic. The results were compared with HPV data obtained by a method using modified filter in situ hybridization and TS-PCR in which the PCR was mainly used to confirm HPV positivity. A substantially higher HPV prevalence rate was found by direct GP/TS-PCR strategy. The results indicate that GP/TS-PCR is a rapid, sensitive, and reliable detection method for HPV in cervical scrapes. The easy performance on crude cell suspensions makes this strategy applicable for large HPV-screening programs.

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Selected References

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  1. Campion M. J., McCance D. J., Cuzick J., Singer A. Progressive potential of mild cervical atypia: prospective cytological, colposcopic, and virological study. Lancet. 1986 Aug 2;2(8501):237–240. doi: 10.1016/s0140-6736(86)92067-2. [DOI] [PubMed] [Google Scholar]
  2. Clewley J. P. The polymerase chain reaction, a review of the practical limitations for human immunodeficiency virus diagnosis. J Virol Methods. 1989 Aug;25(2):179–187. doi: 10.1016/0166-0934(89)90031-1. [DOI] [PubMed] [Google Scholar]
  3. Gregoire L., Arella M., Campione-Piccardo J., Lancaster W. D. Amplification of human papillomavirus DNA sequences by using conserved primers. J Clin Microbiol. 1989 Dec;27(12):2660–2665. doi: 10.1128/jcm.27.12.2660-2665.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Jiwa N. M., Van Gemert G. W., Raap A. K., Van de Rijke F. M., Mulder A., Lens P. F., Salimans M. M., Zwaan F. E., Van Dorp W., Van der Ploeg M. Rapid detection of human cytomegalovirus DNA in peripheral blood leukocytes of viremic transplant recipients by the polymerase chain reaction. Transplantation. 1989 Jul;48(1):72–76. doi: 10.1097/00007890-198907000-00017. [DOI] [PubMed] [Google Scholar]
  5. Kogan S. C., Doherty M., Gitschier J. An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A. N Engl J Med. 1987 Oct 15;317(16):985–990. doi: 10.1056/NEJM198710153171603. [DOI] [PubMed] [Google Scholar]
  6. Koss L. G. The Papanicolaou test for cervical cancer detection. A triumph and a tragedy. JAMA. 1989 Feb 3;261(5):737–743. [PubMed] [Google Scholar]
  7. Li H. H., Gyllensten U. B., Cui X. F., Saiki R. K., Erlich H. A., Arnheim N. Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature. 1988 Sep 29;335(6189):414–417. doi: 10.1038/335414a0. [DOI] [PubMed] [Google Scholar]
  8. Melchers W. J., Herbrink P., Walboomers J. M., Meijer C. J., vd Drift H., Lindeman J., Quint W. G. Optimization of human papillomavirus genotype detection in cervical scrapes by a modified filter in situ hybridization test. J Clin Microbiol. 1989 Jan;27(1):106–110. doi: 10.1128/jcm.27.1.106-110.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Melchers W., van den Brule A., Walboomers J., de Bruin M., Burger M., Herbrink P., Meijer C., Lindeman J., Quint W. Increased detection rate of human papillomavirus in cervical scrapes by the polymerase chain reaction as compared to modified FISH and southern-blot analysis. J Med Virol. 1989 Apr;27(4):329–335. doi: 10.1002/jmv.1890270413. [DOI] [PubMed] [Google Scholar]
  10. Mincheva A., Gissmann L., zur Hausen H. Chromosomal integration sites of human papillomavirus DNA in three cervical cancer cell lines mapped by in situ hybridization. Med Microbiol Immunol. 1987;176(5):245–256. doi: 10.1007/BF00190531. [DOI] [PubMed] [Google Scholar]
  11. Mullink H., Walboomers J. M., Tadema T. M., Jansen D. J., Meijer C. J. Combined immuno- and non-radioactive hybridocytochemistry on cells and tissue sections: influence of fixation, enzyme pre-treatment, and choice of chromogen on detection of antigen and DNA sequences. J Histochem Cytochem. 1989 May;37(5):603–609. doi: 10.1177/37.5.2467928. [DOI] [PubMed] [Google Scholar]
  12. Shibata D. K., Arnheim N., Martin W. J. Detection of human papilloma virus in paraffin-embedded tissue using the polymerase chain reaction. J Exp Med. 1988 Jan 1;167(1):225–230. doi: 10.1084/jem.167.1.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Snijders P. J., van den Brule A. J., Schrijnemakers H. F., Snow G., Meijer C. J., Walboomers J. M. The use of general primers in the polymerase chain reaction permits the detection of a broad spectrum of human papillomavirus genotypes. J Gen Virol. 1990 Jan;71(Pt 1):173–181. doi: 10.1099/0022-1317-71-1-173. [DOI] [PubMed] [Google Scholar]
  14. Wagner D., Ikenberg H., Boehm N., Gissmann L. Identification of human papillomavirus in cervical swabs by deoxyribonucleic acid in situ hybridization. Obstet Gynecol. 1984 Dec;64(6):767–772. [PubMed] [Google Scholar]
  15. van den Brule A. J., Claas E. C., du Maine M., Melchers W. J., Helmerhorst T., Quint W. G., Lindeman J., Meijer C. J., Walboomers J. M. Use of anticontamination primers in the polymerase chain reaction for the detection of human papilloma virus genotypes in cervical scrapes and biopsies. J Med Virol. 1989 Sep;29(1):20–27. doi: 10.1002/jmv.1890290105. [DOI] [PubMed] [Google Scholar]
  16. van den Brule A. J., Snijders P. J., Gordijn R. L., Bleker O. P., Meijer C. J., Walboomers J. M. General primer-mediated polymerase chain reaction permits the detection of sequenced and still unsequenced human papillomavirus genotypes in cervical scrapes and carcinomas. Int J Cancer. 1990 Apr 15;45(4):644–649. doi: 10.1002/ijc.2910450412. [DOI] [PubMed] [Google Scholar]
  17. zur Hausen H. Papillomavirus in anogenital cancer: the dilemma of epidemiologic approaches. J Natl Cancer Inst. 1989 Nov 15;81(22):1680–1682. doi: 10.1093/jnci/81.22.1680. [DOI] [PubMed] [Google Scholar]
  18. zur Hausen H. Papillomaviruses in anogenital cancer as a model to understand the role of viruses in human cancers. Cancer Res. 1989 Sep 1;49(17):4677–4681. [PubMed] [Google Scholar]

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