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Journal of Clinical Microbiology, April 1999, p. 1030-1034, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Human Papillomavirus (HPV) DNA Copy Number Is Dependent on Grade of Cervical Disease and HPV Type

David C. Swan,1,* Ruth Ann Tucker,1 Guillermo Tortolero-Luna,2 Michele Follen Mitchell,2 Louise Wideroff,3 Elizabeth R. Unger,1 Rosane A. Nisenbaum,1 William C. Reeves,1 and Joseph P. Icenogle1

National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Atlanta, Georgia 303331; Department of Gynecologic Oncology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 770302; and Division of Cancer Control and Population Science, National Cancer Institute, Bethesda, Maryland 20892-73443

Received 4 December 1998/Returned for modification 8 January 1999/Accepted 22 January 1999

The association between human papillomavirus (HPV) DNA copy number and cervical disease was investigated. Viral DNA copy number for the most common high-risk HPV types in cervical cancer (types 16, 18, 31, and 45) was determined in cervical cytobrush specimens from 149 women with high-grade cervical intraepithelial neoplasias (CIN II-CIN III), 176 with low-grade CIN (CIN I), and 270 with normal cytology. Quantitative, PCR-based fluorescent assays for each of the HPV genotypes and for the beta -globin gene were used. The amount of cellular DNA increased significantly with increasing disease; thus, HPV was expressed as copies per microgram of cellular DNA. The assay had a dynamic range of >107, allowing documentation for the first time of the wide range of HPV copy numbers seen in clinical specimens. Median HPV DNA copy number varied by more than 104 among the viral types. HPV16 was present in the highest copy number; over 55% of HPV16-positive samples contained more than 108 copies/µg. Median copy number for HPV16 showed dramatic increases with increasing epithelial abnormality, an effect not seen with the other HPV types. HPV16 increased from a median of 2.2 × 107 in patients with normal cytology, to 4.1 × 107 in CIN I patients, to 1.3 × 109 copies/µg in CIN II-III patients. Even when stratified by cervical disease and viral type, the range of viral DNA copies per microgram of cellular DNA was quite large, precluding setting a clinically significant cutoff value for "high" copy numbers predictive of disease. This study suggests that the clinical usefulness of HPV quantitation requires reassessment and is assay dependent.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, Mail Stop G-18, 1600 Clifton Rd., Atlanta, GA 30333. Phone: (404) 639-1300. Fax: (404) 639-0049. E-mail: dcs1{at}cdc.gov.

Journal of Clinical Microbiology, April 1999, p. 1030-1034, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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