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. 1991 May;82(5):524–531. doi: 10.1111/j.1349-7006.1991.tb01882.x

Detection and Typing of Multiple Genital Human Papillomaviruses by DNA Amplification with Consensus Primers

Hiroyuki Yoshikawa 1, Takashi Kawana 3, Kyoko Kitagawa 1, Masahiko Mizuno 1, Hiroshi Yoshikura 2, Aikichi Iwamoto 2
PMCID: PMC5918477  PMID: 1648051

Abstract

Many types of human papillomavirus (HPV) are associated with genital lesions. In order to develop simple and sensitive diagnostic procedures for HPV infection, we took advantage of the polymerase chain reaction (PCR). We compared the published nucleotide sequences of the LI region from six genital HPV types and designed a pair of consensus primers for LI region. The PCR with the consensus primers for LI region (Ll‐PCR) could amplify at least nine genital HPV types, 6,11, 16, 18, 31, 33, 42, 52 and 58, and the amplified HPV DNA could be typed by subsequent restriction mapping. Ll‐PCR was compared to Southern blot analysis and also to the consensus primer‐mediated PCR for E6 region (E6‐PCR) described before. Although both our PCR systems are nonradioactive, the sensitivity in detecting HPV DNA was even better than that obtained by using Southern blot analysis. By means of the PCR systems we detected HPV DNA in 100% of cervical condylomas (10/10), 92% of cervical intraepithelial neoplasias (33/36) and 96% of invasive cervical carcinomas (53/55), while we detected HPV DNA in 12% of normal cervices (12/102).

Keywords: Human papillomavirus, Carvical cancer, Polymerase chain reaction

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REFERENCES

  • 1. ) Tsunokawa , Y. , Takebe , N. , Kasamatsu , T. , Terada , M. and Sugimura , T.Transforming activity of human papillomavirus type 16 DNA sequences in a cervical cancer . Proc. Natl Acad. Sci. USA , 83 , 2200 – 2203 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. ) Yasumoto , S. , Burkhardt , A. L. , Doniger , J. and DiPaolo , J. A.Human papillomavirus type 16 DNA‐induced malignant transformation of NIH3T3 cells . J. Viral , 57 , 572 – 577 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. ) Kanda , T. , Watanabe , S. and Yoshiike , K.Human papillomavirus type 16 transformation of rat 3Y1 cells , Jpn. J. Cancer Res. , 78 , 103 – 108 ( 1987. ). [PubMed] [Google Scholar]
  • 4. ) Pirisi , L. , Yasumoto , S. , Feller , M. , Doniger , J. and DiPaolo , J. A.Transformation of human fibroblasts and keratinocytes with human papillomavirus type 16 DNA . J. Virol , 61 , 1061 – 1066 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5. ) Kanda , T. , Furuno , A. and Yoshiike , K.Human papillomavirus type 16 open reading frame E7 encodes a transforming gene for rat 3YI cells . J. Virol. , 62 , 610 – 613 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6. ) Schwarz , E. , Freese , U. K. , Gissmann , L. , Mayer , W. , Roggenbuck , B. , Stremlau , A. and zur Hausen , H.Structure and transcription of human papillomavirus sequences in cervical carcinoma cells . Nature , 314 , 111 – 114 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Pater , M. M. and Pater , A.Human papillomavirus types 16 and 18 sequences in carcinoma cell lines of the cervix . Virology , 145 , 313 – 318 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Matsukura , T. , Kanda , T. , Furuno , A. , Yoshikawa , H. , Kawana , T. and Yoshiike , K.Cloning of monomeric human papillomavirus type 16 DNA integrated within cell DNA from a cervical carcinoma . J. Virol , 58 , 979 – 982 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. ) Smotkin , D. and Wettstein , F. O.Transcription of human papillomavirus type 16 early genes in a cervical cancer and a cancer‐derived cell line and identification of the E7 protein . Proc. Natl. Acad. Sci. USA , 83 , 4680 – 4684 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10. ) Seedorf , K. , Oltersdorf , T. , Kraemmer , G. and Roewekamp , W.Identification of early proteins of the human papilloma viruses type 16 (HPV 16) and type 18 (HPV 18) in cervical carcinoma cells . EMBO J. , 6 , 139 – 144 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. ) Munoz , N. , Bosch , X. and Kaldor , J. M.Does human papillomavirus cause cervical cancer? The state of the epidemiological evidence . Br. J. Cancer , 57 , 1 – 5 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. ) Reeves , W. C. , Rawls , W. E. and Brinton , L. A.Epidemiology of genital papillomaviruses and cervical cancer . Rev. Infect. Dis. , 11 , 426 – 439 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Lorincz , A. T. , Lancaster , W. D. and 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. , 58 , 225 – 229 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. ) Beaudenon , S. , Kremsdorf , D. , Croissant , O. , Jablonska , S. , Wain‐Hobson , S. and Orth , G.A novel type of human papillomavirus associated with genital neoplasias . Nature , 321 , 246 – 249 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Lorincz , A. T. , Quinn , A. P. , Lancaster , W. D. and Temple , G. F.A new type of papillomavirus associated with cancer of the uterine cervix . Virology , 159 , 187 – 190 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) Beaudenon , S. , Kremsdorf , D. , Obalek , S. , Jablonska , S. , Pehau‐Arnaudet , G. , Croissant , O. and Orth , G.Plurality of genital human papillomaviruses: characterization of two new types with distinct biological properties . Virology , 161 , 374 – 384 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 17. ) Shimoda , K. , Lorincz , A. T. , Temple , G. F. and Lancaster , W. D.Human papillomavirus type 52: a new virus associated with cervical neoplasia . J. Gen. Virol. , 69 , 2925 – 2928 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) 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]
  • 19. ) 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]
  • 20. ) Saiki , R. K. , Scharf , S. , Faloona , F. , Mullis , K. B. , Horn , G. T. , Erlich , H. A. and Arnheim , N.Enzymatic amplification of β‐globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia . Science , 230 , 1350 – 1354 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Saiki , R. K , Gelfand , D. H. , Stofrel , S. , Scharf , S. J , Higuchi , R , Horn , G. T , Mullis , K. B. and Ehrlich , H. A.Primer‐directed enzymatic amplification of DNA with a thermostable DNA polymerase . Science , 239 , 487 – 491 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Shibata , D. K. , Arnheim , N. and Martin , W. J.Detection of human papilloma virus in paraffin‐embedded tissue using the polymerase chain reaction . J. Exp. Med. , 167 , 225 – 230 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Shimada , M. , Fukushima , M. , Mukai , H. , Kato , L. , 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]
  • 24. ) Schwarz , E. , Duerst , M. , Demankowski , C. , Lattermann , O. , Zech , R. , Wolfsperger , E. , Suhai , S. and zur Hausen , H.DNA sequence and genome organization of genital human papillomavirus type 6b . EMBO J. , 12 , 2341 – 2348 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. ) Dartmann , K. , Schwarz , E. , Gissmann , L. and zur Hausen , H.The nucleotide sequence and genome organization of human papillomavirus type 11 . Virology , 151 , 124 – 130 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Seedorf , K. , Kraemmer , G. , Duerst , M. , Suhai , S. and Roewekamp , W. G.Human papillomavirus type 16 DNA sequence . Virology , 145 , 181 – 185 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Cole , S. T. and Danos , O.Nucleotide sequence and comparative analysis of the human papillomavirus type 18 genome . J. Mol Biol , 193 , 599 – 608 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Goldsborough , M. D. , DiSilvestre , D. , Temple , G. F. and Lorincz , A. T.Nucleotide sequence of human papillomavirus type 31: a cervical neoplasia‐associated virus . Virology , 171 , 306 – 311 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) 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]
  • 30. ) 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]
  • 31. ) 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]
  • 32. ) van den Brule , J. C. A. , Snijders , P. J. F. , Gordijn , R. L. J. , Bicker , O. P. , Meijer , C. J. L. M. and Walboomers , J. M. M.General primer‐mediated polymerase chain reaction permits the detection of sequenced and still unsequenced human papillomavirus genotypes in cervical scrapes and carcinomas , Int. J. Cancer , 45 , 644 – 649 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Yoshikawa , H. , Kawana , T. , Kitagawa , K. , Mizuno , M. , Yoshikura , H. and Iwamoto , A.Amplification and typing of multiple cervical cancer‐associated human papillomavirus DNAs using a single pair of primers . Int. J. Cancer , 45 , 990 – 992 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 34. ) Yoshikawa , H. , Matsukura , T. , Yamamoto , E. , Kawana , T. , Mizuno , M. and Yoshiike , K.Occurrence of human papillomavirus type 16 and 18 DNA in cervical carcinomas from Japan: age of patients and histological type of carcinomas . Jpn. J. Cancer Res. , 76 , 667 – 671 ( 1985. ). [PubMed] [Google Scholar]
  • 35. ) Pfister , H. and Fuchs , P. G , Papillomaviruses: particles, genome organisation and proteins . In “ Papillomaviruses and Human Disease ”, ed. Syrjaenen K. , Gissmann L. and Koss L. G. , pp. 1 – 18 ( 1987. ). Springer‐Verlag; , Berlin . [Google Scholar]
  • 36. ) 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. , 17 , 173 – 181 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 37. ) Reid , R. , Greenberg , M. , Jenson , B. , Husain , M. , Willett , J. , Daoud , Y. , Temple , G. , Stanhope , C. R. , Sherman , A. I. , Phibbs , G. D. and Lorincz , A. T.Sexually transmitted papillomaviral infections . Am. J. Obstet. Gynecol , 156 , 212 – 222 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 38. ) Yokota , H. , Yoshikawa , H. , Shiromizu , K. , Kawana , T. and Mizuno , M , Detection of human papillomavirus type 6/11, 16 and 18 in exfoliated cells from the uterine cervices of Japanese women with and without lesions . Jpn. J. Cancer Res. , 18 , 896 – 901 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39. ) Koutsky , L. A. , Galloway , D. A. and Holmes , K. K.Epidemiology of genital human papillomavirus infection . Epidemiol. Rev. , 10 , 122 – 163 ( 1988. ). [DOI] [PubMed] [Google Scholar]

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