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Journal of Clinical Microbiology, October 2007, p. 3245-3250, Vol. 45, No. 10
0095-1137/07/$08.00+0     doi:10.1128/JCM.00216-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Human Papillomavirus (HPV) Genotyping Using Paired Exfoliated Cervicovaginal Cells and Paraffin-Embedded Tissues To Highlight Difficulties in Attributing HPV Types to Specific Lesions{triangledown}

Patti E. Gravitt,1* Leen Jan van Doorn,2 Wim Quint,2 Mark Schiffman,3 Allan Hildesheim,3 Andrew G. Glass,4 Brenda B. Rush,4 Jared Hellman,5 Mark E. Sherman,3 Robert D. Burk,6 and Sophia S. Wang3

Departments of Epidemiology and Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland,1 DDL Diagnostic Laboratories, Voorburg, The Netherlands,2 Hormonal and Reproductive Epidemiology Branch, Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20852-7234,3 Kaiser Permanente, Portland, Oregon,4 Information Management Services, Silver Spring, Maryland,5 Albert Einstein College of Medicine, Bronx, New York6

Received 28 January 2007/ Returned for modification 21 March 2007/ Accepted 3 August 2007

Defining type-specific human papillomavirus (HPV) infections within cervical tissues is important for understanding the pathogenesis of cervical neoplasia and assessing the effectiveness of prophylactic vaccines with limited type-specific spectra. We compared HPV DNA-testing results from 146 matched exfoliated-cell and formalin-fixed-tissue specimens collected by cervicovaginal lavage (CVL) within 90 days of each other from women with histologically confirmed cervical intraepithelial lesions (CIN). The CVL specimens were HPV typed using a MY09/11 L1 consensus primer PCR method followed by dot blot hybridization. The tissue specimens were HPV typed using an SPF10 line probe assay HPV detection system. Of the 146 specimen pairs with evidence of CIN in the tissue, 91.8% were positive for one or more HPV types in both the tissue and cellular specimens. Tissue sections were more likely to be HPV negative (P < 0.01). Typing directly from tissue sections resolved multiple infections detected in exfoliated cells to a single HPV type in only 46.9% of cases. Combined use of both specimen types to attribute lesions to HPV type 16 (HPV-16) and/or -18 led to 43.1% attributed to HPV-16 and/or -18 by both specimen types and 19.9% attributed to HPV-16 and/or -18 by one, but not both, specimen types. Unambiguous attribution of cervical lesions to a single, specific HPV type remains a difficult proposition. Use of multiple specimen types or the development of highly sensitive and robust in situ hybridization HPV-testing methods to evaluate the certainty of attribution of lesions to HPV types might provide insights in future efforts, including HPV vaccine trials.

* Corresponding author. Mailing address: Department of Epidemiology, The Johns Hopkins Bloomberg School of Public Health, 615 N. Wolfe St., Room E6535, Baltimore, MD 21205. Phone: (443) 287-6179. Fax: (410) 955-1383. E-mail: pgravitt{at}jhsph.edu

{triangledown} Published ahead of print on 15 August 2007.

Journal of Clinical Microbiology, October 2007, p. 3245-3250, Vol. 45, No. 10
0095-1137/07/$08.00+0     doi:10.1128/JCM.00216-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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