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Department of Pathology, Section of Molecular Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
Sequence analysis of human papillomavirus (HPV) general primer GP5/6 mediated PCR products revealed the presence of short highly conserved sequences adjacent to the 3' ends of both primers. Part of these sequences was used to elongate GP5 and GP6 at their 3' ends to generate the primers GP5+ and GP6+, respectively. Compared with the GP5/6 PCR, GP5+/6+ specific PCR on 22 cloned mucosotropic HPVs revealed an improved HPV detection, reflected by a 10- to 100-fold higher sensitivity and a markedly increased signal to background ratio, especially at the gel level. As determined on purified DNA, the sensitivity of this GP5+/6+ based assay was at the femtogram level for those HPV genotypes which match strongly with the primers (e.g. HPV-16) and at the picogram level for HPV types (e.g. HPV-39 and -51) having four or more mismatches with one or both primers. Application of both methods on 264 cervical scrapes of a cohort of women participating in a prospective follow-up study revealed an increase of total HPV positivity from 39% (GP5/6 PCR) to 43% (GP5+/6+PCR) of the scrapes. Additional HPV typing by PCR specific for the HPV-6, -11, -16, -18, -31 and -33 revealed that all GP5+/6+ PCR positive cases which were negative by GP5/6 PCR (n = 12) contained HPV types different from these six types. These data indicate that the GP5+/6+ PCR method provides an increased detection level mainly of uncommon, apparently poorly matched HPV types in cervical scrapes and most likely in the enlargement of the spectrum of HPVs detectable by this assay.
* Author for correspondence. Fax +31 20 4442964.
Received 12 September 1994;
accepted 29 November 1994.
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A. M. Kaufmann, P. L. Stern, E. M. Rankin, H. Sommer, V. Nuessler, A. Schneider, M. Adams, T. S. Onon, T. Bauknecht, U. Wagner, et al. Safety and Immunogenicity of TA-HPV, a Recombinant Vaccinia Virus Expressing Modified Human Papillomavirus (HPV)-16 and HPV-18 E6 and E7 Genes, in Women with Progressive Cervical Cancer Clin. Cancer Res., December 1, 2002; 8(12): 3676 - 3685. [Abstract] [Full Text] [PDF]
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T Matsha, R Erasmus, A B Kafuko, D Mugwanya, A Stepien, and M I Parker Human papillomavirus associated with oesophageal cancer J. Clin. Pathol., August 1, 2002; 55(8): 587 - 590. [Abstract] [Full Text] [PDF]
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M A E Nobbenhuis, T J M Helmerhorst, A J C van den Brule, L Rozendaal, L H Jaspars, F J Voorhorst, R H M Verheijen, and C J L M Meijer Primary screening for high risk HPV by home obtained cervicovaginal lavage is an alternative screening tool for unscreened women J. Clin. Pathol., June 1, 2002; 55(6): 435 - 439. [Abstract] [Full Text] [PDF]
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S Strauss, P Sastry, C Sonnex, S Edwards, and J Gray Contamination of environmental surfaces by genital human papillomaviruses Sex Transm Inf, April 1, 2002; 78(2): 135 - 138. [Abstract] [Full Text] [PDF]
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F X Bosch, A Lorincz, N Munoz, C J L M Meijer, and K V Shah The causal relation between human papillomavirus and cervical cancer J. Clin. Pathol., April 1, 2002; 55(4): 244 - 265. [Abstract] [Full Text] [PDF]
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S. E. Strome, A. Savva, A. E. Brissett, B. S. Gostout, J. Lewis, A. C. Clayton, R. McGovern, A. L. Weaver, D. Persing, and J. L. Kasperbauer Squamous Cell Carcinoma of the Tonsils: A Molecular Analysis of HPV Associations Clin. Cancer Res., April 1, 2002; 8(4): 1093 - 1100. [Abstract] [Full Text] [PDF]
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A. J. C. van den Brule, R. Pol, N. Fransen-Daalmeijer, L. M. Schouls, C. J. L. M. Meijer, and P. J. F. Snijders GP5+/6+ PCR followed by Reverse Line Blot Analysis Enables Rapid and High-Throughput Identification of Human Papillomavirus Genotypes J. Clin. Microbiol., March 1, 2002; 40(3): 779 - 787. [Abstract] [Full Text] [PDF]
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F. Coutlee, P. Gravitt, J. Kornegay, C. Hankins, H. Richardson, N. Lapointe, H. Voyer, and E. Franco Use of PGMY Primers in L1 Consensus PCR Improves Detection of Human Papillomavirus DNA in Genital Samples J. Clin. Microbiol., March 1, 2002; 40(3): 902 - 907. [Abstract] [Full Text] [PDF]
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 S. Bayo, F X. Bosch, S. de Sanjose, N. Munoz, A. L. Combita, P. Coursaget, M. Diaz, A. Dolo, A. J. van den Brule, and C. J. Meijer Risk factors of invasive cervical cancer in Mali Int. J. Epidemiol., February 1, 2002; 31(1): 202 - 209. [Abstract] [Full Text] [PDF]
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A. Touze, S. de Sanjose, P. Coursaget, M. R. Almirall, V. Palacio, C. J. L. M. Meijer, J. Kornegay, and F. X. Bosch Prevalence of Anti-Human Papillomavirus Type 16, 18, 31, and 58 Virus-Like Particles in Women in the General Population and in Prostitutes J. Clin. Microbiol., December 1, 2001; 39(12): 4344 - 4348. [Abstract] [Full Text] [PDF]
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K. Szuhai, E. Sandhaus, S. M. Kolkman-Uljee, M. Lemaitre, J.-C. Truffert, R. W. Dirks, H. J. Tanke, G. J. Fleuren, E. Schuuring, and A. K. Raap A Novel Strategy for Human Papillomavirus Detection and Genotyping with SybrGreen and Molecular Beacon Polymerase Chain Reaction Am. J. Pathol., November 1, 2001; 159(5): 1651 - 1660. [Abstract] [Full Text] [PDF]
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D. A. Payne, M. V. Straten, D. Carrasco, and S. K. Tyring Molecular Diagnosis of Skin-Associated Infectious Agents Arch Dermatol, November 1, 2001; 137(11): 1497 - 1502. [Abstract] [Full Text] [PDF]
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M. A. Rubin, B. Kleter, M. Zhou, G. Ayala, A. L. Cubilla, W. G. V. Quint, and E. C. Pirog Detection and Typing of Human Papillomavirus DNA in Penile Carcinoma : Evidence for Multiple Independent Pathways of Penile Carcinogenesis Am. J. Pathol., October 1, 2001; 159(4): 1211 - 1218. [Abstract] [Full Text]
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M. Terai and R. D. Burk Characterization of a novel genital human papillomavirus by overlapping PCR: candHPV86 identified in cervicovaginal cells of a woman with cervical neoplasia J. Gen. Virol., September 1, 2001; 82(9): 2035 - 2040. [Abstract] [Full Text] [PDF]
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S. J. Klug, R. Wilmotte, C. Santos, M. Almonte, R. Herrero, I. Guerrero, E. Caceres, D. Peixoto-Guimaraes, G. Lenoir, P. Hainaut, et al. TP53 Polymorphism, HPV Infection, and Risk of Cervical Cancer Cancer Epidemiol. Biomarkers Prev., September 1, 2001; 10(9): 1009 - 1012. [Abstract] [Full Text] [PDF]
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S. Lanham, A. Herbert, A. Basarab, and P. Watt Detection of Cervical Infections in Colposcopy Clinic Patients J. Clin. Microbiol., August 1, 2001; 39(8): 2946 - 2950. [Abstract] [Full Text] [PDF]
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N. C. Sjo, S. Heegaard, J. U. Prause, C. von Buchwald, and H. Lindeberg Human papillomavirus in conjunctival papilloma Br J Ophthalmol, July 1, 2001; 85(7): 785 - 787. [Abstract] [Full Text] [PDF]
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 L.-M. AALTONEN, E. AUVINEN, J. DILLNER, M. LEHTINEN, J. PAAVONEN, H. RIHKANEN, and A. VAHERI Poor antibody response against human papillomavirus in adult-onset laryngeal papillomatosis J. Med. Microbiol., May 1, 2001; 50(5): 468 - 471. [Abstract] [Full Text] [PDF]
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P K S Chan, D P C Chan, K-F To, M Y Yu, J L K Cheung, and A F Cheng Evaluation of extraction methods from paraffin wax embedded tissues for PCR amplification of human and viral DNA J. Clin. Pathol., May 1, 2001; 54(5): 401 - 403. [Abstract] [Full Text] [PDF]
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J. Mork, A. K. Lie, E. Glattre, S. Clark, G. Hallmans, E. Jellum, P. Koskela, B. Moller, E. Pukkala, J. T. Schiller, et al. Human Papillomavirus Infection as a Risk Factor for Squamous-Cell Carcinoma of the Head and Neck N. Engl. J. Med., April 12, 2001; 344(15): 1125 - 1131. [Abstract] [Full Text] [PDF]
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S Lanham, A Herbert, and P Watt HPV detection and measurement of HPV-16, telomerase, and survivin transcripts in colposcopy clinic patients J. Clin. Pathol., April 1, 2001; 54(4): 304 - 308. [Abstract] [Full Text] [PDF]
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 H A Cubie, A L Seagar, E McGoogan, J Whitehead, A Brass, M J Arends, and M W Whitley Rapid real time PCR to distinguish between high risk human papillomavirus types 16 and 18 Mol. Pathol., February 1, 2001; 54(1): 24 - 29. [Abstract] [Full Text]
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E.-S. Abdel-Hady, P. Martin-Hirsch, M. Duggan-Keen, P. L. Stern, J. V. Moore, G. Corbitt, H. C. Kitchener, and I. N. Hampson Immunological and Viral Factors Associated with the Response of Vulval Intraepithelial Neoplasia to Photodynamic Therapy Cancer Res., January 1, 2001; 61(1): 192 - 196. [Abstract] [Full Text]
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E. C. Pirog, B. Kleter, S. Olgac, P. Bobkiewicz, J. Lindeman, W. G. V. Quint, R. M. Richart, and C. Isacson Prevalence of Human Papillomavirus DNA in Different Histological Subtypes of Cervical Adenocarcinoma Am. J. Pathol., October 1, 2000; 157(4): 1055 - 1062. [Abstract] [Full Text] [PDF]
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L Rozendaal, J Westerga, J C van der Linden, J M M Walboomers, F J Voorhorst, E K J Risse, M E Boon, and C J L M Meijer PCR based high risk HPV testing is superior to neural network based screening for predicting incident CIN III in women with normal cytology and borderline changes J. Clin. Pathol., August 1, 2000; 53(8): 606 - 611. [Abstract] [Full Text] [PDF]
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W. M. Lin, E. A. Michalopulos, N. Dhurander, P. C. Cheng, W. Robinson, R. Ashfaq, R. L. Coleman, and C. Y. Muller Allelic Loss and Microsatellite Alterations of Chromosome 3p14.2 Are More Frequent in Recurrent Cervical Dysplasias Clin. Cancer Res., April 1, 2000; 6(4): 1410 - 1414. [Abstract] [Full Text]
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A. Petter, K. Heim, M. Guger, A. Ciresa-Kö nig, N. Christensen, M. Sarcletti, U. Wieland, H. Pfister, R. Zangerle, and R. Höpfl Specific serum IgG, IgM and IgA antibodies to human papillomavirus types 6, 11, 16, 18 and 31 virus-like particles in human immunodeficiency virus-seropositive women J. Gen. Virol., March 1, 2000; 81(3): 701 - 708. [Abstract] [Full Text]
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J. H. Nelson, G. A. Hawkins, K. Edlund, M. Evander, L. Kjellberg, G. Wadell, J. Dillner, T. Gerasimova, A. L. Coker, L. Pirisi, et al. A Novel and Rapid PCR-Based Method for Genotyping Human Papillomaviruses in Clinical Samples J. Clin. Microbiol., February 1, 2000; 38(2): 688 - 695. [Abstract] [Full Text]
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M. van Duin, P. J. F. Snijders, M. T. M. Vossen, E. Klaassen, F. Voorhorst, R. H. M. Verheijen, T. J. Helmerhorst, C. J. L. M. Meijer, and J. M. M. Walboomers Analysis of human papillomavirus type 16 E6 variants in relation to p53 codon 72 polymorphism genotypes in cervical carcinogenesis J. Gen. Virol., February 1, 2000; 81(2): 317 - 325. [Abstract] [Full Text]
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P. E. Gravitt, C. L. Peyton, T. Q. Alessi, C. M. Wheeler, F. Coutlée, A. Hildesheim, M. H. Schiffman, D. R. Scott, and R. J. Apple Improved Amplification of Genital Human Papillomaviruses J. Clin. Microbiol., January 1, 2000; 38(1): 357 - 361. [Abstract] [Full Text]
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K.-L. Wallin, F. Wiklund, T. Angstrom, F. Bergman, U. Stendahl, G. Wadell, G. Hallmans, and J. Dillner Type-Specific Persistence of Human Papillomavirus DNA before the Development of Invasive Cervical Cancer N. Engl. J. Med., November 25, 1999; 341(22): 1633 - 1638. [Abstract] [Full Text] [PDF]
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L. A. Koopman, K. Szuhai, J. D. H. van Eendenburg, V. Bezrookove, G. G. Kenter, E. Schuuring, H. Tanke, and G. Jan Fleuren Recurrent Integration of Human Papillomaviruses 16, 45, and 67 Near Translocation Breakpoints in New Cervical Cancer Cell Lines Cancer Res., November 1, 1999; 59(21): 5615 - 5624. [Abstract] [Full Text] [PDF]
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W. J. G. Melchers, J. M. J .E. Bakkers, J. Wang, P. C. M. de Wilde, H. Boonstra, W. G. V. Quint, and A. G. J. M. Hanselaar Short Fragment Polymerase Chain Reaction Reverse Hybridization Line Probe Assay to Detect and Genotype a Broad Spectrum of Human Papillomavirus Types : Clinical Evaluation and Follow-Up Am. J. Pathol., November 1, 1999; 155(5): 1473 - 1478. [Abstract] [Full Text] [PDF]
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O. Forslund, A. Antonsson, P. Nordin, B. Stenquist, and B. Göran Hansson A broad range of human papillomavirus types detected with a general PCR method suitable for analysis of cutaneous tumours and normal skin J. Gen. Virol., September 1, 1999; 80(9): 2437 - 2443. [Abstract] [Full Text]
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H. J. Bontkes, T. D. de Gruijl, A. Bijl, R. H. M. Verheijen, C. J. L. M. Meijer, R. J. Scheper, P. L. Stern, J. E. Burns, N. J. Maitland, and J. M. M. Walboomers Human papillomavirus type 16 E2-specific T-helper lymphocyte responses in patients with cervical intraepithelial neoplasia J. Gen. Virol., September 1, 1999; 80(9): 2453 - 2459. [Abstract] [Full Text]
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B. Kleter, L.-J. van Doorn, L. Schrauwen, A. Molijn, S. Sastrowijoto, J. ter Schegget, J. Lindeman, B. ter Harmsel, M. Burger, and W. Quint Development and Clinical Evaluation of a Highly Sensitive PCR-Reverse Hybridization Line Probe Assay for Detection and Identification of Anogenital Human Papillomavirus J. Clin. Microbiol., August 1, 1999; 37(8): 2508 - 2517. [Abstract] [Full Text]
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A. Josefsson, K. Livak, and U. Gyllensten Detection and Quantitation of Human Papillomavirus by Using the Fluorescent 5' Exonuclease Assay J. Clin. Microbiol., March 1, 1999; 37(3): 490 - 496. [Abstract] [Full Text]
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