1887

Abstract

Sequence analysis of human papillomavirus (HPV) general primer GP5/6 mediated PCR products revealed the presence of short highly conserved sequences adjacent to the 3′ ends of both primers. Part of these sequences was used to elongate GP5 and GP6 at their 3′ ends to generate the primers GP5+ and GP6+, respectively. Compared with the GP5/6 PCR, GP5+/6+ specific PCR on 22 cloned mucosotropic HPVs revealed an improved HPV detection, reflected by a 10- to 100-fold higher sensitivity and a markedly increased signal to background ratio, especially at the gel level. As determined on purified DNA, the sensitivity of this GP5+/6+ based assay was at the femtogram level for those HPV genotypes which match strongly with the primers (e.g. HPV-16) and at the picogram level for HPV types (e.g. HPV-39 and -51) having four or more mismatches with one or both primers. Application of both methods on 264 cervical scrapes of a cohort of women participating in a prospective follow-up study revealed an increase of total HPV positivity from 39% (GP5/6 PCR) to 43% (GP5+/6+PCR) of the scrapes. Additional HPV typing by PCR specific for the HPV-6, -11, -16, -18, -31 and -33 revealed that all GP5+/6+ PCR positive cases which were negative by GP5/6 PCR ( = 12) contained HPV types different from these six types. These data indicate that the GP5+/6+ PCR method provides an increased detection level mainly of uncommon, apparently poorly matched HPV types in cervical scrapes and most likely in the enlargement of the spectrum of HPVs detectable by this assay.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-76-4-1057
1995-04-01
2022-01-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/76/4/JV0760041057.html?itemId=/content/journal/jgv/10.1099/0022-1317-76-4-1057&mimeType=html&fmt=ahah

References

  1. DeRoda Husman A. M., Walboomers J. M. M., Meijer C. J. L. M., Risse E. K. J., Schipper M. E. I., Helmerhorst T. M., Bleker O. P., Delius H., Van den Brule A. J. C., Snijders P. J. F. 1994; Analysis of cytomorphologically abnormal cervical scrapes for the presence of 27 mucosotropic human papillomavirus genotypes using polymerase chain reaction. International Journal of Cancer 56:802–806
    [Google Scholar]
  2. de Villiers E.-M. 1989; Heterogeneity in the human papillomavirus group. Journal of Virology 63:4898–4903
    [Google Scholar]
  3. Evander M., Wadell G. 1991; A general primer pair for amplification and detection of genital human papillomavirus types. Journal of Virological Methods 31:239–250
    [Google Scholar]
  4. Grégoire L., Arella M., Campione-Piccardo J., Lancaster W. D. 1989; Amplification of human papillomavirus DNA sequences by using conserved primers. Journal of Clinical Microbiology 27:2660–2665
    [Google Scholar]
  5. Mack D. H., Sninsky J. J. 1988; A sensitive method for the identification of uncharacterized viruses related to known virus groups: hepadnavirus model system. Proceedings of the National Academy of Sciences, USA 85:6977–6981
    [Google Scholar]
  6. Manos M. M., Ting Y., Wright D. K., Lewis A. J., Broker T. R., Wolinsky S. M. 1989; Use of polymerase chain reaction amplification for the detection of genital human papillomaviruses. Cancer Cells 7:209–214
    [Google Scholar]
  7. Matsukura T., Sugase M. 1990; Molecular cloning of a novel human papillomavirus (type 58) from an invasive cervical carcinoma. Virology 177:833–836
    [Google Scholar]
  8. Newton C. R., Graham A., Heptinstall L. E., Powell S. J., Summers C., Kalsheker N., Smith J. C., Markham A. F. 1989; Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acids Research 17:2503–2516
    [Google Scholar]
  9. Reuter S., Delius H., Kahn T., Hofmann B., zur Hausen H., Schwartz E. 1991; Characterization of a novel human papillomavirus DNA in the cervical carcinoma cell line ME 180. Journal of Virology 65:5564–5568
    [Google Scholar]
  10. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. 1985; Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle-cell anemia. Science 230:1350–1354
    [Google Scholar]
  11. Smits H. L., Tieben L. M., Tjong-A-Hung S. P., Jebbink M. F., Minnaar R. P., Jansen C. L., ter Schegget J. 1992; Detection and typing of human papillomaviruses present in fixed and stained archival cervical smears by a consensus polymerase chain reaction and direct sequence analysis allow the identification of a broad spectrum of human papillomavirus types. Journal of General Virology 73:3263–3268
    [Google Scholar]
  12. Snuders P. J. F., Van den Brule A. J. C., Schrijnemakers H. F. J., Snow G., Meijer C. J. L. M., Walboomers J. M. M. 1990; The use of general primers in the polymerase chain reaction permits the detection of a broad spectrum of human papillomavirus genotypes. Journal of General Virology 71:173–181
    [Google Scholar]
  13. Sommer R., Tautz D. 1989; Minimal homology requirements for PCR primers. Nucleic Acids Research 17:6749
    [Google Scholar]
  14. Tawheed A. R., Beaudenon S., Favre M., Orth G. 1991; Characterization of human papillomavirus type 66 from an invasive carcinoma of the uterine cervix. Journal of Clinical Microbiology 29:2656–2660
    [Google Scholar]
  15. Van den Brule A. J. C., Snuders P. J. F., Gordijn R. L. J., Bleker O. P., Meijer C. J. L. M., Walboomers J. M. M. 1990a; General primer polymerase chain reaction permits the detection of sequenced and still unsequenced human papillomavirus genotypes in cervical scrapes and carcinomas. International Journal of Cancer 45:644–649
    [Google Scholar]
  16. Van den Brule A. J. C., Meijer C. J. L. M., Bakels V., Kenemans P., Walboomers J. M. M. 1990b; Rapid detection of human papillomavirus in cervical scrapes by combined general primer-mediated and type-specific polymerase chain reaction. Journal of Clinical Microbiology 28:2739–2743
    [Google Scholar]
  17. Van den Brule A. J. C., Walboomers J. M. M., Du Maine M., Kenemans P., Meijer C. J. L. M. 1991; Difference in prevalence of human papillomavirus genotypes incytomorphologically normal cervical smears is associated with a history of cervical intraepithelial neoplasia. International Journal of Cancer 48:404–408
    [Google Scholar]
  18. Van den Brule A. J. C., Snijders P. J. F., Raaphorst P. M. C., Schrijnemakers H. F. J., Delius H., Gissmann L., Meijer C. J. L. M., Walboomers J. M. M. 1992; General primer polymerase chain reaction in combination with sequence analysis for identification of potentially novel human papillomavirus genotypes in cervical lesions. Journal of Clinical Microbiology 30:1716–1721
    [Google Scholar]
  19. Van den Brule A. J. C., Snijders P. J. F., Meuer C. J. L. M., Walboomers J. M. M. 1993; PCR based detection of genital HPV genotypes: an update and future perspectives. Papillomavirus Reports 4:95
    [Google Scholar]
  20. Vooijs G. P. 1987; De advisering bij afwijkende bevindingen van cytologisch onderzoek van de cervix uteri. Nederlands Tijdschrift voor Geneeskunde 131:1662–1663
    [Google Scholar]
  21. Walboomers J. M. M., Melkert P. W. J., Van den Brule A. J. C., Snijders P. J. F., Meijer C. J. L. M. 1992; The polymerase chain reaction for human papillomavirus screening in diagnostic cytopathology of the cervix. In Diagnostic Molecular Pathology: a Practical Approach Vol II pp 153–172 Edited by Herrington C. S., McGee J. O.-D. Oxford: Oxford University Press;
    [Google Scholar]
  22. Zur Hausen H. 1991; Human papillomaviruses in the pathogenesis of anogenital cancer. Virology 184:9–13
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-76-4-1057
Loading
/content/journal/jgv/10.1099/0022-1317-76-4-1057
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error