Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Mar;33(3):690–695. doi: 10.1128/jcm.33.3.690-695.1995

Nested PCR approach for detection and typing of epidermodysplasia verruciformis-associated human papillomavirus types in cutaneous cancers from renal transplant recipients.

R J Berkhout 1, L M Tieben 1, H L Smits 1, J N Bavinck 1, B J Vermeer 1, J ter Schegget 1
PMCID: PMC228015  PMID: 7751378

Abstract

The epidermodysplasia verruciformis (EV)-associated human papillomaviruses (HPVs) constitute a group of HPV genotypes isolated mostly from the cutaneous lesions of patients with the genetic disorder of EV. Broad-spectrum detection of EV HPVs in cutaneous lesions of non-EV patients was previously difficult because no EV HPV consensus PCR was available. We describe a nested PCR that enables the detection of all known EV HPV types at relatively low-copy-number levels. The deduced sequences of a 92-amino-acid stretch of the L1 open reading frames of all types are shown for convenient typing. The technique proved very valuable in viral studies of skin cancers from renal transplant recipients. A high prevalence (81%) of EV HPV types was found in skin cancer biopsies. A wide spectrum of EV HPV types that differed from HPV-5 and -8 was found to be involved. The technique also proved useful in detecting potentially novel EV HPV types in skin cancers. The relationship of these new types to known HPV types is demonstrated by phylogenetic tree analysis.

Full Text

The Full Text of this article is available as a PDF (225.8 KB).

Selected References

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

  1. Barr B. B., Benton E. C., McLaren K., Bunney M. H., Smith I. W., Blessing K., Hunter J. A. Human papilloma virus infection and skin cancer in renal allograft recipients. Lancet. 1989 Jan 21;1(8630):124–129. doi: 10.1016/s0140-6736(89)91143-4. [DOI] [PubMed] [Google Scholar]
  2. Bavinck J. N., Gissmann L., Claas F. H., Van der Woude F. J., Persijn G. G., Ter Schegget J., Vermeer B. J., Jochmus I., Müller M., Steger G. Relation between skin cancer, humoral responses to human papillomaviruses, and HLA class II molecules in renal transplant recipients. J Immunol. 1993 Aug 1;151(3):1579–1586. [PubMed] [Google Scholar]
  3. Bernard H. U., Chan S. Y., Delius H. Evolution of papillomaviruses. Curr Top Microbiol Immunol. 1994;186:33–54. doi: 10.1007/978-3-642-78487-3_3. [DOI] [PubMed] [Google Scholar]
  4. Boom R., Sol C. J., Salimans M. M., Jansen C. L., Wertheim-van Dillen P. M., van der Noordaa J. Rapid and simple method for purification of nucleic acids. J Clin Microbiol. 1990 Mar;28(3):495–503. doi: 10.1128/jcm.28.3.495-503.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bouwes Bavinck J. N., Vermeer B. J., van der Woude F. J., Vandenbroucke J. P., Schreuder G. M., Thorogood J., Persijn G. G., Claas F. H. Relation between skin cancer and HLA antigens in renal-transplant recipients. N Engl J Med. 1991 Sep 19;325(12):843–848. doi: 10.1056/NEJM199109193251203. [DOI] [PubMed] [Google Scholar]
  6. Bouwes Bavinck J. N., Vermeer B. J., van der Woude F. J., Vandenbroucke J. P., Schreuder G. M., Thorogood J., Persijn G. G., Claas F. H. Relation between skin cancer and HLA antigens in renal-transplant recipients. N Engl J Med. 1991 Sep 19;325(12):843–848. doi: 10.1056/NEJM199109193251203. [DOI] [PubMed] [Google Scholar]
  7. Chan S. Y., Bernard H. U., Ong C. K., Chan S. P., Hofmann B., Delius H. Phylogenetic analysis of 48 papillomavirus types and 28 subtypes and variants: a showcase for the molecular evolution of DNA viruses. J Virol. 1992 Oct;66(10):5714–5725. doi: 10.1128/jvi.66.10.5714-5725.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chou Q., Russell M., Birch D. E., Raymond J., Bloch W. Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res. 1992 Apr 11;20(7):1717–1723. doi: 10.1093/nar/20.7.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deau M. C., Favre M., Orth G. Genetic heterogeneity among human papillomaviruses (HPV) associated with epidermodysplasia verruciformis: evidence for multiple allelic forms of HPV5 and HPV8 E6 genes. Virology. 1991 Oct;184(2):492–503. doi: 10.1016/0042-6822(91)90419-c. [DOI] [PubMed] [Google Scholar]
  10. Delius H., Hofmann B. Primer-directed sequencing of human papillomavirus types. Curr Top Microbiol Immunol. 1994;186:13–31. doi: 10.1007/978-3-642-78487-3_2. [DOI] [PubMed] [Google Scholar]
  11. Dopazo J., Sobrino F. A computer program for the design of PCR primers for diagnosis of highly variable genomes. J Virol Methods. 1993 Feb;41(2):157–165. doi: 10.1016/0166-0934(93)90123-9. [DOI] [PubMed] [Google Scholar]
  12. Dyall-Smith D., Trowell H., Mark A., Dyall-Smith M. Cutaneous squamous cell carcinomas and papillomaviruses in renal transplant recipients: a clinical and molecular biological study. J Dermatol Sci. 1991 May;2(3):139–146. doi: 10.1016/0923-1811(91)90059-7. [DOI] [PubMed] [Google Scholar]
  13. Ensser A., Pfister H. Epidermodysplasia verruciformis associated human papillomaviruses present a subgenus-specific organization of the regulatory genome region. Nucleic Acids Res. 1990 Jul 11;18(13):3919–3922. doi: 10.1093/nar/18.13.3919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Favre M., Obalek S., Jablonska S., Orth G. Human papillomavirus (HPV) type 50, a type associated with epidermodysplasia verruciformis (EV) and only weakly related to other EV-specific HPVs. J Virol. 1989 Nov;63(11):4910–4910. doi: 10.1128/jvi.63.11.4910-.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Favre M., Obalek S., Jablonska S., Orth G. Human papillomavirus type 49, a type isolated from flat warts of renal transplant patients. J Virol. 1989 Nov;63(11):4909–4909. doi: 10.1128/jvi.63.11.4909-.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fuchs P. G., Iftner T., Weninger J., Pfister H. Epidermodysplasia verruciformis-associated human papillomavirus 8: genomic sequence and comparative analysis. J Virol. 1986 May;58(2):626–634. doi: 10.1128/jvi.58.2.626-634.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gassenmaier A., Fuchs P., Schell H., Pfister H. Papillomavirus DNA in warts of immunosuppressed renal allograft recipients. Arch Dermatol Res. 1986;278(3):219–223. doi: 10.1007/BF00412927. [DOI] [PubMed] [Google Scholar]
  18. Gassenmaier A., Lammel M., Pfister H. Molecular cloning and characterization of the DNAs of human papillomaviruses 19, 20, and 25 from a patient with epidermodysplasia verruciformis. J Virol. 1984 Dec;52(3):1019–1023. doi: 10.1128/jvi.52.3.1019-1023.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grimmel M., de Villiers E. M., Neumann C., Pawlita M., zur Hausen H. Characterization of a new human papillomavirus (HPV 41) from disseminated warts and detection of its DNA in some skin carcinomas. Int J Cancer. 1988 Jan 15;41(1):5–9. doi: 10.1002/ijc.2910410103. [DOI] [PubMed] [Google Scholar]
  20. Hartevelt M. M., Bavinck J. N., Kootte A. M., Vermeer B. J., Vandenbroucke J. P. Incidence of skin cancer after renal transplantation in The Netherlands. Transplantation. 1990 Mar;49(3):506–509. doi: 10.1097/00007890-199003000-00006. [DOI] [PubMed] [Google Scholar]
  21. Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
  22. Hoxtell E. O., Mandel J. S., Murray S. S., Schuman L. M., Goltz R. W. Incidence of skin carcinoma after renal transplantation. Arch Dermatol. 1977 Apr;113(4):436–438. [PubMed] [Google Scholar]
  23. Jablonska S., Majewski S. Epidermodysplasia verruciformis: immunological and clinical aspects. Curr Top Microbiol Immunol. 1994;186:157–175. doi: 10.1007/978-3-642-78487-3_9. [DOI] [PubMed] [Google Scholar]
  24. Kawashima M., Favre M., Jablonska S., Obalek S., Orth G. Characterization of a new type of human papillomavirus (HPV) related to HPV5 from a case of actinic keratosis. Virology. 1986 Oct 30;154(2):389–394. doi: 10.1016/0042-6822(86)90464-2. [DOI] [PubMed] [Google Scholar]
  25. Kawashima M., Favre M., Obalek S., Jablonska S., Orth G. Premalignant lesions and cancers of the skin in the general population: evaluation of the role of human papillomaviruses. J Invest Dermatol. 1990 Nov;95(5):537–542. doi: 10.1111/1523-1747.ep12504887. [DOI] [PubMed] [Google Scholar]
  26. Kiyono T., Adachi A., Ishibashi M. Genome organization and taxonomic position of human papillomavirus type 47 inferred from its DNA sequence. Virology. 1990 Jul;177(1):401–405. doi: 10.1016/0042-6822(90)90500-q. [DOI] [PubMed] [Google Scholar]
  27. Kremsdorf D., Favre M., Jablonska S., Obalek S., Rueda L. A., Lutzner M. A., Blanchet-Bardon C., Van Voorst Vader P. C., Orth G. Molecular cloning and characterization of the genomes of nine newly recognized human papillomavirus types associated with epidermodysplasia verruciformis. J Virol. 1984 Dec;52(3):1013–1018. doi: 10.1128/jvi.52.3.1013-1018.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kremsdorf D., Jablonska S., Favre M., Orth G. Biochemical characterization of two types of human papillomaviruses associated with epidermodysplasia verruciformis. J Virol. 1982 Aug;43(2):436–447. doi: 10.1128/jvi.43.2.436-447.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kremsdorf D., Jablonska S., Favre M., Orth G. Human papillomaviruses associated with epidermodysplasia verruciformis. II. Molecular cloning and biochemical characterization of human papillomavirus 3a, 8, 10, and 12 genomes. J Virol. 1983 Nov;48(2):340–351. doi: 10.1128/jvi.48.2.340-351.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lutzner M., Croissant O., Ducasse M. F., Kreis H., Crosnier J., Orth G. A potentially oncogenic human papillomavirus (HPV-5) found in two renal allograft recipients. J Invest Dermatol. 1980 Oct;75(4):353–356. doi: 10.1111/1523-1747.ep12531131. [DOI] [PubMed] [Google Scholar]
  31. Müller M., Kelly G., Fiedler M., Gissmann L. Human papillomavirus type 48. J Virol. 1989 Nov;63(11):4907–4908. doi: 10.1128/jvi.63.11.4907-4908.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Obalek S., Favre M., Szymanczyk J., Misiewicz J., Jablonska S., Orth G. Human papillomavirus (HPV) types specific of epidermodysplasia verruciformis detected in warts induced by HPV3 or HPV3-related types in immunosuppressed patients. J Invest Dermatol. 1992 Jun;98(6):936–941. doi: 10.1111/1523-1747.ep12460892. [DOI] [PubMed] [Google Scholar]
  33. Pfister H., Gassenmaier A., Fuchs P. G. Demonstration of human papillomavirus DNA in two keratoacanthomas. Arch Dermatol Res. 1986;278(3):243–246. doi: 10.1007/BF00412933. [DOI] [PubMed] [Google Scholar]
  34. Pfister H. Human papillomaviruses and skin cancer. Semin Cancer Biol. 1992 Oct;3(5):263–271. [PubMed] [Google Scholar]
  35. Rübben A., Krones R., Schwetschenau B., Grussendorf-Conen E. I. Common warts from immunocompetent patients show the same distribution of human papillomavirus types as common warts from immunocompromised patients. Br J Dermatol. 1993 Mar;128(3):264–270. doi: 10.1111/j.1365-2133.1993.tb00169.x. [DOI] [PubMed] [Google Scholar]
  36. Rüdlinger R., Grob R. Papillomavirus infection and skin cancer in renal allograft recipients. Lancet. 1989 May 20;1(8647):1132–1133. doi: 10.1016/s0140-6736(89)92403-3. [DOI] [PubMed] [Google Scholar]
  37. Saiki R. K., Chang C. A., Levenson C. H., Warren T. C., Boehm C. D., Kazazian H. H., Jr, Erlich H. A. Diagnosis of sickle cell anemia and beta-thalassemia with enzymatically amplified DNA and nonradioactive allele-specific oligonucleotide probes. N Engl J Med. 1988 Sep 1;319(9):537–541. doi: 10.1056/NEJM198809013190903. [DOI] [PubMed] [Google Scholar]
  38. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  39. Scheurlen W., Gissmann L., Gross G., zur Hausen H. Molecular cloning of two new HPV types (HPV 37 and HPV 38) from a keratoacanthoma and a malignant melanoma. Int J Cancer. 1986 Apr 15;37(4):505–510. doi: 10.1002/ijc.2910370406. [DOI] [PubMed] [Google Scholar]
  40. Stark L. A., Arends M. J., McLaren K. M., Benton E. C., Shahidullah H., Hunter J. A., Bird C. C. Prevalence of human papillomavirus DNA in cutaneous neoplasms from renal allograft recipients supports a possible viral role in tumour promotion. Br J Cancer. 1994 Feb;69(2):222–229. doi: 10.1038/bjc.1994.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tieben L. M., ter Schegget J., Minnaar R. P., Bouwes Bavinck J. N., Berkhout R. J., Vermeer B. J., Jebbink M. F., Smits H. L. 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. doi: 10.1016/0166-0934(93)90038-s. [DOI] [PubMed] [Google Scholar]
  42. Van Ranst M., Kaplan J. B., Burk R. D. Phylogenetic classification of human papillomaviruses: correlation with clinical manifestations. J Gen Virol. 1992 Oct;73(Pt 10):2653–2660. doi: 10.1099/0022-1317-73-10-2653. [DOI] [PubMed] [Google Scholar]
  43. Van der Leest R. J., Zachow K. R., Ostrow R. S., Bender M., Pass F., Faras A. J. Human papillomavirus heterogeneity in 36 renal transplant recipients. Arch Dermatol. 1987 Mar;123(3):354–357. doi: 10.1001/archderm.123.3.354. [DOI] [PubMed] [Google Scholar]
  44. Yabe Y., Sakai A., Hitsumoto T., Kato H., Ogura H. A subtype of human papillomavirus 5 (HPV-5b) and its subgenomic segment amplified in a carcinoma: nucleotide sequences and genomic organizations. Virology. 1991 Aug;183(2):793–798. doi: 10.1016/0042-6822(91)91013-7. [DOI] [PubMed] [Google Scholar]
  45. al-Bekairi A. M., Qureshi S., Chaudhry M. A., Shah A. H. Uric acid as an inhibitor of cyclophosphamide-induced micronuclei in mice. Mutat Res. 1991 Feb;262(2):115–118. doi: 10.1016/0165-7992(91)90116-l. [DOI] [PubMed] [Google Scholar]
  46. de Villiers E. M. Heterogeneity of the human papillomavirus group. J Virol. 1989 Nov;63(11):4898–4903. doi: 10.1128/jvi.63.11.4898-4903.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. de Villiers E. M. Human pathogenic papillomavirus types: an update. Curr Top Microbiol Immunol. 1994;186:1–12. doi: 10.1007/978-3-642-78487-3_1. [DOI] [PubMed] [Google Scholar]
  48. von Knebel Doeberitz M. Papillomaviruses in human disease: Part I. Pathogenesis and epidemiology of human papillomavirus infections. Eur J Med. 1992 Nov;1(7):415–423. [PubMed] [Google Scholar]

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

RESOURCES