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Journal of Clinical Microbiology, March 2002, p. 902-907, Vol. 40, No. 3
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.3.902-907.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Use of PGMY Primers in L1 Consensus PCR Improves Detection of Human Papillomavirus DNA in Genital Samples

François Coutlée,1,2* Patti Gravitt,3,{dagger} Janet Kornegay,3 Catherine Hankins,4,5 Harriet Richardson,4 Normand Lapointe,6 Hélène Voyer,1 The Canadian Women’s HIV Study Group,,{ddagger} and Eduardo Franco4

Laboratoire de Virologie Moléculaire, Centre de Recherche et Département de Microbiologie et Infectiologie, Hôpital Notre-Dame du Centre Hospitalier de l'Université de Montréal,1 Département de Microbiologie et Immunologie, Université de Montréal,2 Department of Epidemiology and Biostatistics, McGill University,4 Direction de la Santé Publique de Montréal-Centre, Institut National de Santé Publique du Québec,5 Centre Maternel et Infantile sur le SIDA, Centre de Recherche de l'Hôpital Sainte-Justine, Hôpital Sainte-Justine, Montréal, Québec, Canada,6 Roche Molecular Systems, Alameda, California3

Received 7 June 2001/ Returned for modification 30 July 2001/ Accepted 21 December 2001

The novel PGMY L1 consensus primer pair is more sensitive than the MY09 and MY11 primer mix for detection and typing with PCR of human papillomavirus (HPV) DNA in genital specimens. We assessed the diagnostic yield of PGMY primers for the detection and typing of HPV by comparing the results obtained with PGMY09/PGMY11 and MY09/MY11/HMB01 on 299 genital samples. Amplicons generated with PGMY primers were typed with the line blot assay (PGMY-line blot), while HPV amplicons obtained with the degenerate primer pool MY09/MY11/HMB01 were detected with type-specific radiolabeled probes in a dot blot assay (standard consensus PCR test). Cervicovaginal lavage samples (N = 272) and cervical scrape samples (N = 27) were tested in parallel with both PCR tests. The PGMY-line blot test detected the presence of HPV DNA more frequently than the standard consensus PCR assay. The concordance for HPV typing between the two assays was 84.3% (214 of 255 samples), for a good kappa value of 0.69. Of the 177 samples containing HPV DNA by at least one method, 40 samples contained at least one HPV type detected only with PGMY-line blot, whereas positivity exclusively with the standard consensus PCR test was found for only 7 samples (P < 0.001). HPV types 45 and 52 were especially more frequently detected with PGMY than MY primers. However, most HPV types were better amplified with PGMY primers, including HPV-16. Samples with discordant results between the two PCR assays more frequently contained multiple HPV types. Studies using PGMY instead of MY primers have the potential to report higher detection rates of HPV infection not only for newer HPV types but also for well-known genital types.

* Corresponding author. Mailing address: Département de Microbiologie et Infectiologie, Hôpital Notre-Dame du Centre Hospitalier de l'Université de Montréal, 1560 Sherbrooke est, Montréal, Québec H2L 4M1, Canada. Phone: 514-890-8000, ext. 25162. Fax: 514-412-7512. E-mail: francois.coutlee{at}ssss.gouv.qc.ca.

{dagger} Present address: Department of Epidemiology, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Md.

{ddagger} The Canadian Women's HIV Study Group includes the following investigators throughout Canada (coinvestigators are marked with an asterisk). Principal investigator, Catherine Hankins. Halifax: Janet Conners,* Rob Grimshaw, David Haase, Lynn Johnston,* Wally Schlech,* Arlo Yuzicappi-Fayant.* Hamilton: Stephen Landis, Fiona Smaill. London: Tom Austin, Ole Hammerberg, Ted Ralph.* Montréal: François Coutlée,* Julian Falutz, Alex Ferenczy, Marina Klein,* Louise Labrecque, Richard Lalonde, John Macleod, Grégoire Noël,* Chantal Perron,* Jean-Pierre Routy, and Emil Toma. Ottawa: Claire Touchie, Garry Victor.* Québec: Louise Coté, Hélène Senay, Sylvie Trottier. Saskatoon: Kurt Williams.* Sherbrooke: Alain Piché.* Sudbury: Roger Sandre. Toronto: Louise Binder,* Donna Keystone, Anne Phillips, Anita Rachlis,* Irving Salit, Cheryl Wagner, Sharon Walmsley.* Vancouver: Paula Braitstein,* David Burdge,* Marianne Harris, Deborah Money,* Julio Montaner.*

Journal of Clinical Microbiology, March 2002, p. 902-907, Vol. 40, No. 3
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.3.902-907.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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