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. 1989 Dec;27(12):2660–2665. doi: 10.1128/jcm.27.12.2660-2665.1989

Amplification of human papillomavirus DNA sequences by using conserved primers.

L Gregoire 1, M Arella 1, J Campione-Piccardo 1, W D Lancaster 1
PMCID: PMC267104  PMID: 2556429

Abstract

The polymerase chain reaction has potential for use in the detection of small amounts of human papillomavirus (HPV) viral nucleic acids present in clinical specimens. However, new HPV types for which no probes exist would remain undetected by using type-specific primers for the polymerase chain reaction before hybridization. Primers corresponding to highly conserved HPV sequences may be useful for detecting low amounts of known HPV DNA as well as new HPV types. Here we analyze a pair of primers derived from conserved sequences within the E1 open reading frame for HPV sequence amplification by using the polymerase chain reaction. The longest perfect homology among HPV sequences is a 12-mer within the first exon of E1M. A region of conserved amino acids coded by the E1 open reading frame allowed the detection of another highly conserved region about 850 base pairs downstream. Two 21-mers derived from these conserved regions were used to amplify sequences from all HPV DNAs used as templates. The amplified DNA was shown to be specific for HPV sequences within the E1 open reading frame. DNA from HPVs whose sequences were not available were amplified by using these two primers. HPV DNA sequences in clinical specimens could also be amplified with the primers.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barnes W., Delgado G., Kurman R. J., Petrilli E. S., Smith D. M., Ahmed S., Lorincz A. T., Temple G. F., Jenson A. B., Lancaster W. D. Possible prognostic significance of human papillomavirus type in cervical cancer. Gynecol Oncol. 1988 Mar;29(3):267–273. doi: 10.1016/0090-8258(88)90225-9. [DOI] [PubMed] [Google Scholar]
  2. Beaudenon S., Kremsdorf D., Croissant O., Jablonska S., Wain-Hobson S., Orth G. A novel type of human papillomavirus associated with genital neoplasias. Nature. 1986 May 15;321(6067):246–249. doi: 10.1038/321246a0. [DOI] [PubMed] [Google Scholar]
  3. Boshart M., Gissmann L., Ikenberg H., Kleinheinz A., Scheurlen W., zur Hausen H. A new type of papillomavirus DNA, its presence in genital cancer biopsies and in cell lines derived from cervical cancer. EMBO J. 1984 May;3(5):1151–1157. doi: 10.1002/j.1460-2075.1984.tb01944.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chien A., Edgar D. B., Trela J. M. Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus. J Bacteriol. 1976 Sep;127(3):1550–1557. doi: 10.1128/jb.127.3.1550-1557.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cole S. T., Danos O. Nucleotide sequence and comparative analysis of the human papillomavirus type 18 genome. Phylogeny of papillomaviruses and repeated structure of the E6 and E7 gene products. J Mol Biol. 1987 Feb 20;193(4):599–608. doi: 10.1016/0022-2836(87)90343-3. [DOI] [PubMed] [Google Scholar]
  6. Cole S. T., Streeck R. E. Genome organization and nucleotide sequence of human papillomavirus type 33, which is associated with cervical cancer. J Virol. 1986 Jun;58(3):991–995. doi: 10.1128/jvi.58.3.991-995.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dartmann K., Schwarz E., Gissmann L., zur Hausen H. The nucleotide sequence and genome organization of human papilloma virus type 11. Virology. 1986 May;151(1):124–130. doi: 10.1016/0042-6822(86)90110-8. [DOI] [PubMed] [Google Scholar]
  8. Dürst M., Gissmann L., Ikenberg H., zur Hausen H. A papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographic regions. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3812–3815. doi: 10.1073/pnas.80.12.3812. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  10. Fuchs P. G., Girardi F., Pfister H. Human papillomavirus 16 DNA in cervical cancers and in lymph nodes of cervical cancer patients: a diagnostic marker for early metastases? Int J Cancer. 1989 Jan 15;43(1):41–44. doi: 10.1002/ijc.2910430110. [DOI] [PubMed] [Google Scholar]
  11. Fuchs P. G., Girardi F., Pfister H. Human papillomavirus DNA in normal, metaplastic, preneoplastic and neoplastic epithelia of the cervix uteri. Int J Cancer. 1988 Jan 15;41(1):41–45. doi: 10.1002/ijc.2910410109. [DOI] [PubMed] [Google Scholar]
  12. Gissmann L., Diehl V., Schultz-Coulon H. J., zur Hausen H. Molecular cloning and characterization of human papilloma virus DNA derived from a laryngeal papilloma. J Virol. 1982 Oct;44(1):393–400. doi: 10.1128/jvi.44.1.393-400.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Heilman C. A., Law M. F., Israel M. A., Howley P. M. Cloning of human papilloma virus genomic DNAs and analysis of homologous polynucleotide sequences. J Virol. 1980 Nov;36(2):395–407. doi: 10.1128/jvi.36.2.395-407.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kurman R. J., Schiffman M. H., Lancaster W. D., Reid R., Jenson A. B., Temple G. F., Lorincz A. T. Analysis of individual human papillomavirus types in cervical neoplasia: a possible role for type 18 in rapid progression. Am J Obstet Gynecol. 1988 Aug;159(2):293–296. doi: 10.1016/s0002-9378(88)80070-x. [DOI] [PubMed] [Google Scholar]
  15. Law M. F., Lancaster W. D., Howley P. M. Conserved polynucleotide sequences among the genomes of papillomaviruses. J Virol. 1979 Oct;32(1):199–207. doi: 10.1128/jvi.32.1.199-207.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lorincz A. T., Lancaster W. D., Temple G. F. Cloning and characterization of the DNA of a new human papillomavirus from a woman with dysplasia of the uterine cervix. J Virol. 1986 Apr;58(1):225–229. doi: 10.1128/jvi.58.1.225-229.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lorincz A. T., Quinn A. P., Lancaster W. D., Temple G. F. A new type of papillomavirus associated with cancer of the uterine cervix. Virology. 1987 Jul;159(1):187–190. doi: 10.1016/0042-6822(87)90366-7. [DOI] [PubMed] [Google Scholar]
  18. Lorincz A. T., Temple G. F., Kurman R. J., Jenson A. B., Lancaster W. D. Oncogenic association of specific human papillomavirus types with cervical neoplasia. J Natl Cancer Inst. 1987 Oct;79(4):671–677. [PubMed] [Google Scholar]
  19. Lörincz A. T., Quinn A. P., Goldsborough M. D., Schmidt B. J., Temple G. F. Cloning and partial DNA sequencing of two new human papillomavirus types associated with condylomas and low-grade cervical neoplasia. J Virol. 1989 Jun;63(6):2829–2834. doi: 10.1128/jvi.63.6.2829-2834.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ohtsuka E., Matsuki S., Ikehara M., Takahashi Y., Matsubara K. An alternative approach to deoxyoligonucleotides as hybridization probes by insertion of deoxyinosine at ambiguous codon positions. J Biol Chem. 1985 Mar 10;260(5):2605–2608. [PubMed] [Google Scholar]
  21. Pfister H. Relationship of papillomaviruses to anogenital cancer. Obstet Gynecol Clin North Am. 1987 Jun;14(2):349–361. [PubMed] [Google Scholar]
  22. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  23. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  24. Schwarz E., Dürst M., Demankowski C., Lattermann O., Zech R., Wolfsperger E., Suhai S., zur Hausen H. DNA sequence and genome organization of genital human papillomavirus type 6b. EMBO J. 1983;2(12):2341–2348. doi: 10.1002/j.1460-2075.1983.tb01744.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Seedorf K., Krämmer G., Dürst M., Suhai S., Röwekamp W. G. Human papillomavirus type 16 DNA sequence. Virology. 1985 Aug;145(1):181–185. doi: 10.1016/0042-6822(85)90214-4. [DOI] [PubMed] [Google Scholar]
  26. Shah K. V., Gissmann L. Experimental evidence on oncogenicity of papillomaviruses. IARC Sci Publ. 1989;(94):105–111. [PubMed] [Google Scholar]
  27. 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]
  28. Shibata D., Martin W. J., Arnheim N. Analysis of DNA sequences in forty-year-old paraffin-embedded thin-tissue sections: a bridge between molecular biology and classical histology. Cancer Res. 1988 Aug 15;48(16):4564–4566. [PubMed] [Google Scholar]
  29. Shimoda K., Lorincz A. T., Temple G. F., Lancaster W. D. Human papillomavirus type 52: a new virus associated with cervical neoplasia. J Gen Virol. 1988 Nov;69(Pt 11):2925–2928. doi: 10.1099/0022-1317-69-11-2925. [DOI] [PubMed] [Google Scholar]
  30. Tidy J., Vousden K. H., Mason P., Farrell P. J. A novel deletion within the upstream regulatory region of episomal human papillomavirus type 16. J Gen Virol. 1989 Apr;70(Pt 4):999–1004. doi: 10.1099/0022-1317-70-4-999. [DOI] [PubMed] [Google Scholar]
  31. Young L. S., Bevan I. S., Johnson M. A., Blomfield P. I., Bromidge T., Maitland N. J., Woodman C. B. The polymerase chain reaction: a new epidemiological tool for investigating cervical human papillomavirus infection. BMJ. 1989 Jan 7;298(6665):14–18. doi: 10.1136/bmj.298.6665.14. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. de Villiers E. M., Gissmann L., zur Hausen H. Molecular cloning of viral DNA from human genital warts. J Virol. 1981 Dec;40(3):932–935. doi: 10.1128/jvi.40.3.932-935.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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