Summary
The detection of oncogenic subtypes of human papillomavirus (HPV) from exfoliated cervical cells collected in liquid-based preservative fluid at the time of the annual Pap test can be accomplished using PCR-based amplification methods. DNA sequence analysis of the L1 open reading frame (ORF) permits determination of the HPV viral subtype. DNA sequence analysis of the E6 and E7 ORFs permits further detection of various HPV genotype variants within a given HPV viral type. These techniques can be readily adopted by most laboratories employing standard molecular biology techniques.
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References
zur Hausen, H. (2000) Papillomaviruses causing cancer: evasion from host-cell control in early events in carcinogenesis. J Natl Cancer Inst 92, 690–698.
Lorincz, A.T., Reid, R., Jenson, A.B., Green-berg, M.D., Lancaster, W., and Kurman, R.J. (1992) Human papillomavirus infection of the cervix: relative risk associations of 15 common anogenital types. Obstet Gynecol 79, 328–337.
Cogliano, V., Baan, R., Straif, K., Grosse, Y., Secretan, B., El Ghissassi, F. (2005) WHO International Agency for Research on Cancer. Carcinogenicity of human papillomaviruses. Lancet Oncol 6, 204.
Chan, S.Y., Bernard, H.U., Ong, C.K., Chan, S.P., Hofmann, B., and Delius, H. (1992) Phylogenetic analysis of 48 papillomavirus types and 28 subtypes and variants: a showcase for the molecular evolution of DNA viruses. J Virol 66, 5714–5725.
Longworth, M.S., and Laimins, L.A. (2004) Pathogenesis of human papillomaviruses in differentiating epithelia. Microbiol Mol Biol Rev 68, 362–372.
Zheng, Z.M., and Baker, C.C. (2006) Papil-lomavirus genome structure, expression, and post-transcriptional regulation. Front Biosci 11, 2286–2302.
Munger, K., Baldwin, A., Edwards, K.M., et al. (2004) Mechanisms of human papillomavirus-induced oncogenesis. J Virol 78, 11451–1160.
Longworth, M.S., and Laimins, L.A. (2004) Pathogenesis of human papillomaviruses in differentiating epithelia. Microbiol Mol Biol Rev 68, 362–372.
Bernard, H.U. (2005) The clinical importance of the nomenclature, evolution and taxonomy of human papillomaviruses. J Clin Virol 32(1), S1–S6.
Villa, L.L., Sichero, L., Rahal, P., Caballero, O., Ferenczy, A., Rohan, T., and Franco, E.L. (2000) Molecular variants of human papillo-mavirus types 16 and 18 preferentially associated with cervical neoplasia. J Gen Virol 81(Pt 12), 2959–2968.
Calleja-Macias, I.E., Kalantari, M., Huh, J., Ortiz-Lopez, R., Rojas-Martinez, A., Gonzalez-Guerrero, J.F., Williamson, A.L., Hagmar, B., Wiley, D.J., Villarreal, L., Bernard, H.U., and Barrera-Saldaña, H.A. (2004) Genomic diversity of human papillomavirus-16, 18, 31, and 35 isolates in a Mexican population and relationship to European, African, and Native American variants. Virology 319, 315–323.
Calleja-Macias, I.E., Villa, L.L., Prado, J.C., Kalantari, M., Allan, B., Williamson, A.L., Cheng, L.P., Collins, R.J., Zuna, R.E., Dunn, S.T., Chu, T.Y., Cubie, H.A., Cuschieri, K., von Knebel-Doeberitz, M., Martins, C.R., Sanchez, G.I., Bosch, F.X., Munoz, N., and Bernard, H.U. (2005) Worldwide genomic diversity of the high-risk human papillomavi-rus types 31, 35, 52, and 58, four close relatives of human papillomavirus type 16. J Virol 79, 13630–13640.
Arias-Pulido, H., Peyton, C.L., Torrez-Mar tínez, N., Anderson, D.N., and Wheeler, C.M.(2005) Human papillomavirus type 18 variant lineages in United States populations characterized by sequence analysis of LCR-E6, E2, and L1 regions. Virology 338, 22–34.
Song, Y.S., Kee, S.H., Kim, J.W., Park, N.H., Kang, S.B., Chang, W.H., and Lee, H.P. (1997) Major sequence variants in E7 gene of human papillomavirus type 16 from cervical cancerous and noncancerous lesions of Korean women. Gynecol Oncol 66, 275–281.
Zehbe, I., Voglino, G., Delius, H., Wilander, E., and Tommasino, M. (1998) Risk of cervical cancer and geographical variations of human papillomavirus 16 E6 polymorphisms. Lancet 352, 1441–1442.
Hsieh, C.H., Tsao, Y.P., Wang, C.H., Han, C.P., Chang, J.L., Lee, J.Y., and Chen, S.L. (2000) Sequence variants and functional analysis of human papillomavirus type 16 E5 gene in clinical specimens. Arch Virol 145, 2273–2284.
Pérez-Gallego, L., Moreno-Bueno, G., Sarrió, D., Suárez, A., Gamallo, C., and Palacios, J. (2001) Human papillomavirus-16 E6 variants in cervical squamous intraepithelial lesions from HIV-negative and HIV-positive women. Am J Clin Pathol 116, 143–148.
Chakrabarti, O., Veeraraghavalu, K., Ter-gaonkar, V., Liu, Y., Androphy, E.J., Stanley, M.A., and Krishna, S. (2004) Human papil-lomavirus type 16 E6 amino acid 83 variants enhance E6-mediated MAPK signaling and differentially regulate tumorigenesis by notch signaling and oncogenic Ras. J Virol 78, 5934–5945.
De Boer, M.A., Peters, L.A., Aziz, M.F., Siregar, B., Cornain, S., Vrede, M.A., Jordanova, E.S., Kolkman-Uljee, S., and Fleuren, G.J. (2004) Human papillomavirus type 16 E6, E7, and L1 variants in cervical cancer in Indonesia, Suriname, and The Netherlands. Gyne-col Oncol 94, 488–494.
De Boer, M.A., Peters, L.A., Aziz, M.F., Siregar, B., Cornain, S., Vrede, M.A., Jordanova, E.S., and Fleuren, G.J. (2005) Human papil-lomavirus type 18 variants: histopathology and E6/E7 polymorphisms in three countries. Int J Cancer 114, 422–425.
Wheeler, C.M., Yamada, T., Hildesheim, A., and Jenison, S.A. (1997) Human papilloma-virus type 16 sequence variants: identification by E6 and L1 lineage-specific hybridization. J Clin Microbiol 35, 11–19.
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Avissar, P., Malinowski, D.P. (2009). Detection and Genotyping Analysis of Human Papillomavirus Isolates from Liquid-Based Cervical Cytology Specimens. In: Kozlov, S.V. (eds) Inflammation and Cancer. Methods in Molecular Biology™, vol 511. Humana Press. https://doi.org/10.1007/978-1-59745-447-6_16
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DOI: https://doi.org/10.1007/978-1-59745-447-6_16
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