General primer-mediated polymerase chain reaction permits the detection of sequenced and still unsequenced human papillomavirus genotypes in cervical scrapes and carcinomas
Corresponding Author
Adriaan J. C. Van Den Brule
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The NetherlandsSearch for more papers by this authorPeter J. F. Snijders
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Search for more papers by this authorRoel L. J. Gordijn
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Search for more papers by this authorOtto P. Bleker
Department of Gynecology, OLVG Hospital, Amsterdam, The Netherlands
Search for more papers by this authorChris J. L. M. Meijer
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Search for more papers by this authorJan M. M. Walboomers
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Search for more papers by this authorCorresponding Author
Adriaan J. C. Van Den Brule
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The NetherlandsSearch for more papers by this authorPeter J. F. Snijders
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Search for more papers by this authorRoel L. J. Gordijn
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Search for more papers by this authorOtto P. Bleker
Department of Gynecology, OLVG Hospital, Amsterdam, The Netherlands
Search for more papers by this authorChris J. L. M. Meijer
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Search for more papers by this authorJan M. M. Walboomers
Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Search for more papers by this authorAbstract
A newly developed general primer-mediated polymerase chain reaction (GP-PCR) was used for the detection of a broad spectrum of Human Papilloma-virus (HPV) genotypes, including unsequenced types, in cytologically normal and abnormal cervical smears and in biopsies of cervical carcinomas. This PCR method used different general primer sets, located in strongly conserved E1 and L1 regions of the HPV genome. Comparison between results of GP-PCR and HPV-type-specific PCR (TS-PCR) revealed an increase in overall HPV prevalence to 25%, 80% and 88% in scrapes with normally, slightly and severely dysplastic cells, respectively. Unsequenced HPV types were detected in 11% of cytologically normal swabs and in up to 30% of scrapes with dysplastic cells. Further characterization showed that unsequenced types concern HPV 13, 30, 31, 45, 51 and some other, possibly unknown HPV types. More than 90% of carcinomas in situ and invasive cervical carcinomas contained HPV. In the latter, only HPV16 and HPV18 were present. HPV16 was most frequently found in both normal and dysplastic cells, the rate being highest in neoplastic tissue. These results indicate that GP-PCR is a powerful approach for detecting as yet uncharacterized HPV types associated with neoplastic transformation of cervical squamous cell epithelium.
References
-
Campion, H. J.,
Cuzick, J.,
McCance, D. J. and
Singer, A.,
Progressive potential of mild cervical atypia: prospective cytological, colposcopic and virological study.
Lancet.
11, 237–240
(1986).
10.1016/S0140-6736(86)92067-2 Google Scholar
- Choo, K. B., Lee, H. H., Pan, C. C., Wu, S. M., Liew, L. N., Cheun, W. F. and Han, S. H., Sequence duplication and internal deletion in the integrated human papillomavirus type 16 genome clones from a cervical carcinoma. J. Virol., 62, 1659–1666 (1988).
- Clewly, J. P., The polymerase chain reaction, a review of the practical limitations for human immunodeficiency virus diagnosis. J. virol. Meth., 25, 179–188 (1989).
- 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. J. mol. Biol., 193, 599–608 (1987).
- Cole, S. T. and Streeck, R., Genome organization and nucleotide sequence of human papillomavirus type 33, which is associated with cervical cancer. J. Virol., 58, 991–995 (1986).
- Dartmann, K., Schwarz, E., Gissmann, L. and Zur Hausen, H., The nucleotide sequence and genome organization of human papilloma virus type 11. Virology, 151, 124–130 (1986).
- De Villiers, E. M., Heterogeneity of the human papillomavirus group. J. Virol., 63, 4898–4903 (1989).
- Gissmann, L., Dürst, M., Oltersdorf, T. and Von Knebel Doeberitz, M., Human papillomaviruses and cervical cancer. Cancer Cells, 5, 275–280 (1987).
- Goldsborough, M. D., Di-Silvestre, 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).
- Grégoire, 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).
- Kahn, T., Schwarz, E. and Zur Hausen, H., Molecular cloning and characterization of the DNA of a new human papillomavirus (HPV30) from a laryngeal carcinoma. Int. J. Cancer, 37, 61–65 (1986).
- Koss, L. G., The Papanicolaou test for cervical cancer detection, a triumph and a tragedy. J. Amer. med. Ass., 261, 737–743 (1989).
- Li, H., Gyllensten, U. B., Cui, X., Saiki, R. K., Erlich, H. A. and Arnheim, N., Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature (Lond.), 335, 414–417 (1988).
- 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).
- Maniatis, T., Fritsch, E. F. and Sambrook, J., Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York (1982).
- 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).
- Melchers, W., Van Den Brule, A., Walboomers, J., De Bruin, M., Herbrink, P., Meijer, C., Lindeman, J. and Quint, W., Increased detection rate of human papillomavirus in cervical scrapes by the polymerase chain reaction as compared to modified FISH and Southern blot analysis. J. med. Virol., 27, 329–335 (1989).
- Naghashfar, Z. S., Rosenshein, N. B., Lorincz, A. T., Buscema, J. and Shah, K. V., Characterization of human papillomavirus type 45, a new type 18-related virus of the genital tract. J. gen. Virol., 68, 3073–3079 (1987).
- Nuovo, G. J., Crum, C. P., De Villiers, E. M., Levine, R. U. and Silverstein, S. J., Isolation of a novel human papillomavirus (HPV51) from a cervical condyloma. J. Virol., 62, 1452–1455 (1988).
- Pfister, H., Hettich, I., Runne, U., Gissmann, L. and Chiff, G. N., Characterization of human papillomavirus type 13 from focal epithelial hyperplasia neck lesions. J. Virol., 47, 363–366 (1983).
- Saiki, R. K., Gelfand, D. H., Stoffel, 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).
- Schwarz, E., Dürst, 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., 2, 2341–3248 (1983).
- 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 (Lond.), 314, 111–114 (1985).
- Seedorf, K., Krämmer, G., Dürst, M., Suhai, S. and Röwekamp, W. G., Human papillomavirus type 16 DNA sequence. Virology, 145, 181–185 (1985).
- 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).
- Tidy, J. A., Vousden, K. H. and Farrell, P. J., Relation between infection with a subtype of HPV16 and cervical neoplasia. Lancet, 1, 1225–1227 (1989).
- Van Den Brule, A. J. C., Claas, H. 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 papillomavirus genotypes in cervical scrapes and biopsies. J. med. Virol., 29, 20–27 (1989).
- Voous, G. P., De advisering bij afwijkende bevindingen van cytologisch onderzoek van de cervix uteri. Ned. Tijdschr. Geneeskd., 131, 1662–1663 (1987).
- Walboomers, J. M. M., Melchers, W. J. G., Mullink, H., Meijer, C. J. L. M., Struyk, A., Quint, W. G. J., Van Der Noordaa, J. and Ter Schegget, J., Sensitivity of in situ detection with biotinylated probes of human papilloma virus type 16 DNA in frozen tissue sections of squamous cell carcinomas of the cervix. Amer. J. Path., 131, 587–594 (1988).
- Wilczynski, S. P., Pearlman, L. and Walker, J., Identification of HPV 16 early genes retained in cervical carcinomas. Virology, 166, 624–627 (1988).
- 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. Brit. med. J., 298, 14–18 (1989).
- 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).
- Zur Hausen, H., Gissmann, L. and Schlehofer, J. R., Viruses in the etiology of human genital cancer. Progr. med. Virol., 30, 170–186 (1984).
- Zur Hausen, H. and Schneider, A., The role of papillomaviruses in human anogenital cancer. In: N. P. Salzman and P. M. Howley (eds.), The papovaviridae–the papillomaviruses, pp. 245–263, Plenum, New York (1987).