Skip to main content
PMC home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1991 Apr;82(4):411–419. doi: 10.1111/j.1349-7006.1991.tb01864.x

Use of Universal and Type‐specific Primers in the Polymerase Chain Reaction for the Detection and Typing of Genital Human Papillomaviruses

Hideo Maid 1, Shigeru Saito 2, Tamotsu Ibaraki 2, Motohiko Ichijo 2, Osanm Yoshie 1,✉
PMCID: PMC5918436  PMID: 1646198

Abstract

By using polymerase chain reaction (PCR), we have developed a system for type‐specific as well as universal detection of genital human papillomaviruses (HPVs). Primers and probes for specific detection of HPV‐16, ‐18 and ‐33 were synthesized from the E7 open reading frame (ORF). They were capable of detecting corresponding HPV types with high specificity and sensitivity. Primers for detection of a broad spectrum of HPV (universal primers) were synthesized from the LI ORF. The universal primers were shown to be capable of amplifying HPV‐6b, ‐11, ‐16, ‐18, ‐33, ‐52b and ‐58. The system was applied to various cervical tissue specimens from Japanese patients. They consisted of 26 normal specimens, 18 from cervical dysplasias and 29 from cervical carcinomas. HPV was detected in none of the normal specimens. On the other hand, many of the specimens from cervical dysplasias and carcinomas were found to be positive for HPV, especially HPV‐16. Except for one, all the specimens which were positive with the type‐specific PCRs were also positive with the universal PCR. Furthermore, substantial numbers of specimens were found to be positive only with the universal PCR. Cloning and sequencing of DNA segments amplified by the universal primers were undertaken to characterize some of the unknown HPVs. Our PCR system may thus be useful for the specific detection of the three major types of oncogenic HPVs and also for the detection of a broad spectrum of HPVs including possibly novel HPV types.

Keywords: Human papillomavirus, Cervical carcinoma, Cervical dysplasia, Polymerase chain reaction

Full Text

The Full Text of this article is available as a PDF (542.5 KB).

REFERENCES

  • 1. ) Pfister , H.Human papillomaviruses and genital cancer . Adv. Cancer Res. , 48 , 113 – 147 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) zur Hausen , H.Papillomaviruses as carcinomaviruses . Adv. Viral Oncol , 8 , 1 – 26 ( 1989. ). [Google Scholar]
  • 3. ) Saito , J. , Yutsudo , M. , Inoue , M , Ueda , G. , Tanizawa , O. and Hakura , A.New human papillomavirus sequences in female genital tumors from Japanese patients . Jpn. J. Cancer Res. 78 , 1081 – 1087 ( 1987. ). [PubMed] [Google Scholar]
  • 4. ) Shibata , D. , Fu , Y. S. , Gupta , J. W. , Shah , K. V. , Arnheim , N. and Martin , W. J.Detection of human papillomavirus in normal and dysplastic tissue by the polymerase chain reaction . Lab. Invest. , 59 , 555 – 559 ( 1988. ). [PubMed] [Google Scholar]
  • 5. ) Young , L. S. , Bevan , I. S. , Johnson , M. A. , Blomfield , P. I. , Bromidge , T. , Maitland , N. J. and Woodman , C. B. J.The polymerase chain reaction: a new epidemiological tool for investigating cervical human papillomavirus infection . Br. Med. J. , 298 , 14 – 18 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6. ) Shimada , M. , Fukushima , M. , Mukai , H. , Kato , I. , Nishikawa , A. and Fujinaga , K.Amplification and specific detection of transforming gene region of human papillomavirus 16, 18 and 33 in cervical carcinoma by means of the polymerase chain reaction . Jpn. J. Cancer Res. , 81 , 1 – 5 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. ) Yokota , H. , Yoshikawa , H. , Shiromizu , K. , Kawana , T. and Mizuno , M.Detection of human papillomavirus types 6/11, 16 and 18 in exfoliated cells from the uterine cervices of Japanese women with and without lesions . Jpn. J. Cancer Res. , 81 , 896 – 901 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8. ) Yajima , H , Noda , T. , de Villiers , E. ‐M. , Yajima , A , Yamamoto , K. , Noda , K. and Ito , Y.Isolation of a new type of human papillomavirus (HPV52b) with a transforming activity from cervical cancer tissue . Cancer Res. , 48 , 7164 – 7172 ( 1988. ). [PubMed] [Google Scholar]
  • 9. ) Matsukura , T. and Sugase , M.Molecular cloning of a novel human papillomavirus (Type 58) from an invasive cervical carcinoma . Virology , 177 , 833 – 836 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) Gregoire , L. , Arella , M. , Campione‐Piccardo , J. and Lancaster , W. D.Amplification of human papillomavirus DNA sequences by using conserved primers . J. Clin. Microbiol , 27 , 2660 – 2665 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. ) Manos , M. M , Ting , Y , Wright , D. K , Lewis , A. J. , Broker , T. R. and Wolinsky , S. M.Use of polymerase chain reaction amplification for the detection of genital human papillomaviruses . Cancer Cells , 7 , 209 – 214 ( 1989. ). [Google Scholar]
  • 12. ) Snijders , P. J. F. , van den Brule , A. J. C. , Schrijnemakers , H. F. J. , Snow , G. , Meijer , C. J. L. M. and Walboomers , J. M. 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 , 71 , 173 – 181 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Maniatis , T. , Fritsch , E. F. and Sambrook , J. “ Molecular Cloning” ( 1982. ). Cold Spring Harbor Laboratory; , New York . [Google Scholar]
  • 14. ) Boshart , M. , Gissmann , L. , Ikenberg , H. , Kleinheinz , A. , Scheurlen , W. and 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. , 3 , 1151 – 1157 ( 1984. . [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. ) Beaudenon , S. , Kremsdorf , D. , Croissant , O. , Jablonska , S. , Wain‐Hobson , S. and Orth , G. Anovel type of human papillomavirus associated with genital neoplasias . Nature , 321 , 246 – 249 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) Heilmann , C. A. , Law , M‐F. , Israel , M. A. and Howley , P. M.Cloning of human papilloma virus genomic DNAs and analysis of homologous polynucleotide sequences , J. Virol. , 36 , 395 – 407 ( 1980. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. ) de Villiers , E‐M. , Gissmann , L. and zur Hausen , H.Molecular cloning of viral DNA from human genital warts . J. Viral , 40 , 932 – 935 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. ) Gissmann , L. , Diehl , V. , Schultz‐Coulon , H‐J. and zur Hausen , H.Molecular cloning and characterization of human papillomavirus DNA derived from a laryngeal papilloma . J. Virol , 44 , 393 – 400 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. ) Dürst , M. , Gissmann , L. , Ikenberg , H. and 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. USA , 80 , 3812 – 3815 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. ) Seedorf , K. , Krämmer , G. , Dürst , M. , Suhai , S. and Röwekamp , W. G.Human papillomavirus type 16 DNA sequence . Virology , 145 , 181 – 185 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Cole , S. T. and 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. / . Mol. Biol. , 193 , 599 – 608 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Cole , S. T. and Streeck , R. E.Genome organization and nucleotide sequence of human papillomavirus type 33, which is associated with cervical cancer . J. Virol. , 58 , 991 – 995 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Saiki , R. K. , Gelfand , D. H. , Stoffel , S. , Scharf , S. J. , Higuchi , R. , Horn , G. L , Mullis , K. B. and Erlich , H. A.Primer‐directed enzymatic amplification of DNA with a thermostable DNA polymerase . Science , 239 , 487 – 491 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 24. ) zur Hausen , H.Papillomaviruses in anogenital cancer as a model to understand the role of viruses in human cancers . Cancer Res. , 49 , 4677 – 4681 ( 1989. ). [PubMed] [Google Scholar]
  • 25. ) Tidy , J. A. , Parry , G. C. N. , Ward , P. , Coleman , D. V. , Peto , J. , Malcolm , A. D. B. and Farrell , P. J.High rate of human papillomavirus type 16 infection in cytologically normal cervices . Lancet , i , 434 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) van den Brule , A. J. C. , Claas , E. C. J. , du Maine , M. , Melchers , W. J. G. , Helmerhorst , T. , Quint , W. G. V. , Lindeman , J. , Meijer , C. J. L. M. and Walboomers , J. M. M.Use of anti‐contamination primers in the polymerase chain reaction for the detection of human papilloma virus genotypes in cervical scrapes and biopsies . J. Med. Virol. , 29 , 20 – 27 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Parton , A.Nucleotide sequence of the HPV16 LI open reading frame . Nucleic Acids Res. , 18 , 3631 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES