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A type‐specific, quantitative assay for the detection of human papillomavirus type 52 (HPV52)
Author(s) -
Onyekwuluje Juanita Marie,
Swan David C,
Tucker Ruth Ann,
Unger Elizabeth R
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.lb248
Subject(s) - multiplex , human papillomavirus , virology , biology , microbiology and biotechnology , medicine , genetics
Human papillomavirus type 52 (HPV52) is a high risk mucosal HPV that is a member of the a‐9 species that includes HPV 16. The potential for HPV vaccines that target HPV 16 to provide cross‐protection against closely related types in the same species increase the importance of unambiguous detection. Unfortunately one of the most widely used commercially available HPV typing assays, the Roche Linear Array based studies detects HPV 52 using the XR probe which cross‐reacts with 33, 35and 58, preventing unambiguous detection of HPV 52 if these types are present. We developed a quantitative multiplex 5′‐exonuclease fluorescent probe real‐time PCR assay for HPV52 and ß‐globin detection and quantitation. The assay specificity was validated using samples evaluated using Roche Linear Array and pyrosequencing of the product. Based on dilution series of synthetic templates, the method is accurate over a large copy number range (from 10 to 10 million copies). We have used the assay successfully on over 1,500 samples collected in PreservCyt, Digene STM, and SurPath media, extracted by a variety of methods. The ability to simultaneously monitor HPV DNA content of the sample has the added advantage of allowing ongoing QC of sample collection and extraction.