1887

Abstract

Human papillomaviruses (HPVs) are aetiological agents for cervical cancer. More than 70 different HPV types that infect genital mucosa have been found. In order to develop a sensitive and specific detection and typing assay, a PCR/direct sequencing approach was used. Two pairs of consensus primers were used for amplification of HPV DNA and the PCR products obtained were analysed by automated sequencing. Sequences were compared with those in GenBank by using the program. In this study, 2916 cytological samples were screened for HPV, as well as for triage. Nine hundred and forty-eight (32.5 %) samples were positive for HPV, of which 134 harboured more than one HPV type. Of the 948 PCR-positive samples, 648 were typed. Thirty-nine different HPV types were identified by sequencing. The two most frequently found HPV types, 16 and 31, together accounted for 36.3 % of the sequences (26.2 and 10.1 %, respectively). This group was followed by HPV types 6 (5.7 %), 18 (5.3 %), 58 (4.5 %), 61 (4.5 %), 53 (4.4 %), 42 (4.3 %) and 51 (4.0 %). All other types were detected at frequencies <4 % and eight types were detected only once. PCR/direct sequencing is a reliable method for routine detection of HPV in cytological samples. The data presented here suggest a complex distribution of HPV types in the population tested. The results accentuate the importance of PCR-based techniques in HPV diagnosis, as hybridization-based methods can only detect a limited number of infections. This method can also be applied easily to the analysis of tissue samples and it therefore also allows type-specific follow-up of women who have been treated for cervical intraepithelial neoplasia.

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2004-02-01
2019-11-17
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References

  1. Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. ( 1997;). Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402.[CrossRef]
    [Google Scholar]
  2. Bauer, H. M., Greer, C. E. & Manos, M. M. ( 1992;). Determination of genital HPV infection using consensus PCR. In Diagnostic Molecular Pathology: a Practical Approach, pp. 131–152. Edited by C. S. Herrington & J. O. McGee. Oxford: Oxford University Press.
  3. Bollmann, R., Méhes, G., Torka, R., Speich, N., Schmitt, C. & Bollmann, M. ( 2003a;). Human papillomavirus typing and DNA ploidy determination of squamous intraepithelial lesions in liquid-based cytologic samples. Cancer Cytopathol 99, 57–62.
    [Google Scholar]
  4. Bollmann, R., Méhes, G., Torka, R., Speich, N., Schmitt, C. & Bollmann, M. ( 2003b;). Determination of features indicating progression in atypical squamous cells with undetermined significance. Cancer Cytopathol 99, 113–117.
    [Google Scholar]
  5. Bosch, F. X., Manos, M. M., Munoz, N. & 7 other authors ( 1995;). Prevalence of human papillomavirus in cervical cancer: a worldwide perspective.International biological study on cervical cancer (IBSCC) Study Group. J Natl Cancer Inst 87, 796–802.[CrossRef]
    [Google Scholar]
  6. Cuzick, J. ( 2000;). Human papillomavirus testing for primary cervical cancer screening. JAMA 283, 108–109.[CrossRef]
    [Google Scholar]
  7. Feoli-Fonseca, J. C., Oligny, L. L., Brochu, P., Simard, P., Falconi, S. & Yotov, W. V. ( 2001;). Human papillomavirus (HPV) study of 691 pathological specimens from Quebec by PCR-direct sequencing approach. J Med Virol 63, 284–292.[CrossRef]
    [Google Scholar]
  8. Jacobs, M. V., Snijders, P. J. F., van den Brule, A. J. C., Helmerhorst, T. J. M., Meijer, C. J. L. M. & Walboomers, J. M. M. ( 1997;). A general primer GP5+/GP6+-mediated PCR-enzyme immunoassay method for rapid detection of 14 high-risk and 6 low-risk human papillomavirus genotypes in cervical scrapings. J Clin Microbiol 35, 791–795.
    [Google Scholar]
  9. Jacobs, M. V., Snijders, P. J. F., Voorhorst, F. J. & 12 other authors ( 1999;). Reliable high risk HPV DNA testing by polymerase chain reaction: an intermethod and intramethod comparison. J Clin Pathol 52, 498–503.[CrossRef]
    [Google Scholar]
  10. Kösel, S., Burggraf, S., Mommsen, J., Engelhardt, W. & Olgemöller, B. ( 2003;). Type-specific detection of human papillomaviruses in a routine laboratory setting –improved sensitivity and specificity of PCR and sequence analysis compared to direct hybridisation. Clin Chem Lab Med 41, 787–791.
    [Google Scholar]
  11. Lorincz, A. T., Reid, R., Jenson, A. B., Greenberg, M. D., Lancaster, W. & Kurman, R. J. ( 1992;). Human papillomavirus infection of the cervix: relative risk associations of 15 common anogenital types. Obstet Gynecol 79, 328–337.[CrossRef]
    [Google Scholar]
  12. Meyer, T., Arndt, R., Beckmann, E. R., Padberg, B., Christophers, E. & Stockfleth, E. ( 2001;). Distribution of HPV 53, HPV 73 and CP8304 in genital epithelial lesions with different grades of dysplasia. Int J Gynecol Cancer 11, 198–204.[CrossRef]
    [Google Scholar]
  13. Muñoz, N., Bosch, F. X., de Sanjosé, S., Herrero, R., Castellsagué, X., Shah, K. V., Snijders, P. J. F. & Meijer, C. J. L. M. ( 2003;). Epidemiologic classification of human papillomavirus types associated with cervical cancer. N Engl J Med 348, 518–527.[CrossRef]
    [Google Scholar]
  14. Nobbenhuis, M. A. E., Meijer, C. J. L. M., van den Brule, A. J. C., Rozendaal, L., Voorhorst, F. J., Risse, E. K. J., Verheijen, R. H. M. & Helmerhorst, T. J. M. ( 2001;). Addition of high-risk HPV testing improves the current guidelines on follow-up after treatment for cervical intraepithelial neoplasia. Br J Cancer 84, 796–801.[CrossRef]
    [Google Scholar]
  15. Szuhai, K., Sandhaus, E., Kolkmann-Uljee, S. M. & 7 other authors ( 2001;). A novel strategy for human papillomavirus detection and genotyping with SybrGreen and molecular beacon polymerase chain reaction. Am J Pathol 159, 1651–1660.[CrossRef]
    [Google Scholar]
  16. van den Brule, A. J. C., Meijer, C. J. L. M., Bakels, V., Kenemans, P. & Walboomers, J. M. ( 1990;). Rapid detection of human papillomavirus in cervical scrapes by combined general primer-mediated and type-specific polymerase chain reaction. J Clin Microbiol 28, 2739–2743.
    [Google Scholar]
  17. van den Brule, A. J. C., Pol, R., Fransen-Daalmeijer, N., Schouls, L. M., Meijer, C. J. L. M. & Snijders, P. J. F. ( 2002;). GP5+/6+ PCR followed by reverse line blot analysis enables rapid and high-throughput identification of human papillomavirus genotypes. J Clin Microbiol 40, 779–787.[CrossRef]
    [Google Scholar]
  18. van Doorn, L.-J., Quint, W., Kleter, B. & 7 other authors ( 2002;). Genotyping of human papillomavirus in liquid cytology cervical specimens by the PGMY line blot assay and the SPF10 line probe assay. J Clin Microbiol 40, 979–983.[CrossRef]
    [Google Scholar]
  19. Walboomers, J. M., Jacobs, M. V., Manos, M. M. & 7 other authors ( 1999;). Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 189, 12–19.[CrossRef]
    [Google Scholar]
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