Search tips
Search criteria 


Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. 1995 April; 33(4): 901–905.
PMCID: PMC228064

Group-specific differentiation between high- and low-risk human papillomavirus genotypes by general primer-mediated PCR and two cocktails of oligonucleotide probes.


In recent years, general primer-mediated PCR assays have been developed to detect a broad spectrum of human papillomavirus (HPV) genotypes. In this study, a procedure enabling a simple group-specific differentiation of high-risk (HPV-16, -18, -31, -33, -35, -39, -45, -51, -52, -54, -56, and -58) and low risk (HPV-6, -11, -34, -40, -42, -43, and 44) HPVs following an HPV general primer-mediated (GP5+/GP6+) PCR is presented. By computer-assisted sequence analysis, oligonucleotides (30-mers) specific for 19 different HPV genotypes were selected from the internal part of the 150-bp GP5+/GP6(+)-amplified region. These oligo probes were tested for specificity in a Southern blot analysis of PCR products derived from the same panel of HPV types. No cross-hybridizations were found. The sensitivities of the oligo probes varied from the femtogram level for the well-amplified HPV types like HPV-16 and -18 to the picogram level for the less-well amplified HPV types like HPV-39 and -51. These sensitivities were reached when the oligo probes were applied both individually and in a cocktail. On the basis of these results, two cocktail oligo probes that enabled a specific and sensitive differentiation between low- and high-risk HPV types were composed.

Full Text

The Full Text of this article is available as a PDF (293K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bergeron C, Barrasso R, Beaudenon S, Flamant P, Croissant O, Orth G. Human papillomaviruses associated with cervical intraepithelial neoplasia. Great diversity and distinct distribution in low- and high-grade lesions. Am J Surg Pathol. 1992 Jul;16(7):641–649. [PubMed]
  • Brinton LA. Epidemiology of cervical cancer--overview. IARC Sci Publ. 1992;(119):3–23. [PubMed]
  • Campion MJ, McCance DJ, Cuzick J, Singer A. Progressive potential of mild cervical atypia: prospective cytological, colposcopic, and virological study. Lancet. 1986 Aug 2;2(8501):237–240. [PubMed]
  • Delius H, Hofmann B. Primer-directed sequencing of human papillomavirus types. Curr Top Microbiol Immunol. 1994;186:13–31. [PubMed]
  • de Roda Husman AM, Walboomers JM, Meijer CJ, Risse EK, Schipper ME, Helmerhorst TM, Bleker OP, Delius H, van den Brule AJ, Snijders PJ. Analysis of cytomorphologically abnormal cervical scrapes for the presence of 27 mucosotropic human papillomavirus genotypes, using polymerase chain reaction. Int J Cancer. 1994 Mar 15;56(6):802–806. [PubMed]
  • de Villiers EM. Heterogeneity of the human papillomavirus group. J Virol. 1989 Nov;63(11):4898–4903. [PMC free article] [PubMed]
  • de Villiers EM. Human pathogenic papillomavirus types: an update. Curr Top Microbiol Immunol. 1994;186:1–12. [PubMed]
  • Eluf-Neto J, Booth M, Muñoz N, Bosch FX, Meijer CJ, Walboomers JM. Human papillomavirus and invasive cervical cancer in Brazil. Br J Cancer. 1994 Jan;69(1):114–119. [PMC free article] [PubMed]
  • Evander M, Wadell G. A general primer pair for amplification and detection of genital human papillomavirus types. J Virol Methods. 1991 Feb-Mar;31(2-3):239–250. [PubMed]
  • Fujinaga Y, Shimada M, Okazawa K, Fukushima M, Kato I, Fujinaga K. Simultaneous detection and typing of genital human papillomavirus DNA using the polymerase chain reaction. J Gen Virol. 1991 May;72(Pt 5):1039–1044. [PubMed]
  • Gaarenstroom KN, Melkert P, Walboomers JMM, Van Den Brule AJC, Van Bommel PFJ, Meyer CJLM, Voorhorst FJ, Kenemans P, Helmerhorst ThJM. Human papillomavirus DNA and genotypes: prognostic factors for progression of cervical intraepithelial neoplasia. Int J Gynecol Cancer. 1994 Mar;4(2):73–78. [PubMed]
  • Gregoire L, Arella M, Campione-Piccardo J, Lancaster WD. Amplification of human papillomavirus DNA sequences by using conserved primers. J Clin Microbiol. 1989 Dec;27(12):2660–2665. [PMC free article] [PubMed]
  • Higgins DG, Sharp PM. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. [PubMed]
  • Kataja V, Syrjänen S, Mäntyjärvi R, Yliskoski M, Saarikoski S, Syrjänen K. Prognostic factors in cervical human papillomavirus infections. Sex Transm Dis. 1992 May-Jun;19(3):154–160. [PubMed]
  • Koutsky LA, Holmes KK, Critchlow CW, Stevens CE, Paavonen J, Beckmann AM, DeRouen TA, Galloway DA, Vernon D, Kiviat NB. A cohort study of the risk of cervical intraepithelial neoplasia grade 2 or 3 in relation to papillomavirus infection. N Engl J Med. 1992 Oct 29;327(18):1272–1278. [PubMed]
  • Lechner MS, Mack DH, Finicle AB, Crook T, Vousden KH, Laimins LA. Human papillomavirus E6 proteins bind p53 in vivo and abrogate p53-mediated repression of transcription. EMBO J. 1992 Aug;11(8):3045–3052. [PubMed]
  • Lorincz AT, Reid R, Jenson AB, Greenberg MD, Lancaster W, Kurman RJ. Human papillomavirus infection of the cervix: relative risk associations of 15 common anogenital types. Obstet Gynecol. 1992 Mar;79(3):328–337. [PubMed]
  • Lungu O, Sun XW, Felix J, Richart RM, Silverstein S, Wright TC., Jr Relationship of human papillomavirus type to grade of cervical intraepithelial neoplasia. JAMA. 1992 May 13;267(18):2493–2496. [PubMed]
  • Cuzick J, Terry G, Ho L, Hollingworth T, Anderson M. Human papillomavirus type 16 in cervical smears as predictor of high-grade cervical intraepithelial neoplasia [corrected]. Lancet. 1992 Apr 18;339(8799):959–960. [PubMed]
  • Münger K, Yee CL, Phelps WC, Pietenpol JA, Moses HL, Howley PM. Biochemical and biological differences between E7 oncoproteins of the high- and low-risk human papillomavirus types are determined by amino-terminal sequences. J Virol. 1991 Jul;65(7):3943–3948. [PMC free article] [PubMed]
  • Muñoz N, Bosch FX, de Sanjosé S, Tafur L, Izarzugaza I, Gili M, Viladiu P, Navarro C, Martos C, Ascunce N, et al. The causal link between human papillomavirus and invasive cervical cancer: a population-based case-control study in Colombia and Spain. Int J Cancer. 1992 Nov 11;52(5):743–749. [PubMed]
  • Schiffman MH, Bauer HM, Hoover RN, Glass AG, Cadell DM, Rush BB, Scott DR, Sherman ME, Kurman RJ, Wacholder S, et al. Epidemiologic evidence showing that human papillomavirus infection causes most cervical intraepithelial neoplasia. J Natl Cancer Inst. 1993 Jun 16;85(12):958–964. [PubMed]
  • Smits HL, Tieben LM, Tjong-A-Hung SP, Jebbink MF, Minnaar RP, Jansen CL, ter Schegget J. 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. J Gen Virol. 1992 Dec;73(Pt 12):3263–3268. [PubMed]
  • Snijders PJ, van den Brule AJ, Schrijnemakers HF, Snow G, Meijer CJ, Walboomers JM. 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. [PubMed]
  • Tawheed AR, Beaudenon S, Favre M, Orth G. Characterization of human papillomavirus type 66 from an invasive carcinoma of the uterine cervix. J Clin Microbiol. 1991 Nov;29(11):2656–2660. [PMC free article] [PubMed]
  • Tieben LM, ter Schegget J, Minnaar RP, Bouwes Bavinck JN, Berkhout RJ, Vermeer BJ, Jebbink MF, Smits HL. Detection of cutaneous and genital HPV types in clinical samples by PCR using consensus primers. J Virol Methods. 1993 May;42(2-3):265–279. [PubMed]
  • van den Brule AJ, Meijer CJ, Bakels V, Kenemans P, Walboomers JM. Rapid detection of human papillomavirus in cervical scrapes by combined general primer-mediated and type-specific polymerase chain reaction. J Clin Microbiol. 1990 Dec;28(12):2739–2743. [PMC free article] [PubMed]
  • van den Brule AJ, Snijders PJ, Raaphorst PM, Schrijnemakers HF, Delius H, Gissmann L, Meijer CJ, Walboomers JM. General primer polymerase chain reaction in combination with sequence analysis for identification of potentially novel human papillomavirus genotypes in cervical lesions. J Clin Microbiol. 1992 Jul;30(7):1716–1721. [PMC free article] [PubMed]
  • Van Den Brule AJ, Walboomers JM, Du Maine M, Kenemans P, Meijer CJ. Difference in prevalence of human papillomavirus genotypes in cytomorphologically normal cervical smears is associated with a history of cervical intraepithelial neoplasia. Int J Cancer. 1991 May 30;48(3):404–408. [PubMed]
  • Van Ranst M, Kaplan JB, Burk RD. Phylogenetic classification of human papillomaviruses: correlation with clinical manifestations. J Gen Virol. 1992 Oct;73(Pt 10):2653–2660. [PubMed]
  • Williamson AL, Rybicki EP. Detection of genital human papillomaviruses by polymerase chain reaction amplification with degenerate nested primers. J Med Virol. 1991 Mar;33(3):165–171. [PubMed]
  • zur Hausen H. Human papillomaviruses in the pathogenesis of anogenital cancer. Virology. 1991 Sep;184(1):9–13. [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)