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High Level of Correlation of Human Papillomavirus-16 DNA Viral Load Estimates Generated by Three Real-time PCR Assays Applied on Genital Specimens

¹ Laboratoire de Virologie Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal; ² Département de Microbiologie et Immunologie, Université de Montréal, Montreal, Quebec, Canada; ³ Departments of Epidemiology, Biostatistics and Occupational Medicine, and Oncology, McGill University; and ⁴ Direction de la Santé Publique de Montréal-Centre, Institut National de Santé Publique du Québec, Montreal, Quebec, Canada; and ⁵ Department of Epidemiology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland

Requests for reprints: François Coutlée, Département de Microbiologie et Infectiologie, Hôpital Notre-Dame du Centre Hospitalier de l'Université de Montréal, 1560 Sherbrooke est, Montreal, Quebec, Canada H2L 4M1. Phone: 514-890-8000, ext. 25162; Fax: 514-412-7512. E-mail: francois.coutlee{at}ssss.gouv.qc.ca

Human papillomavirus-16 (HPV-16) viral load could be a biomarker predictive of the presence of high-grade cervical lesions. Recently, several real-time PCR assays have been developed to accurately measure HPV-16 viral load. However, results from various reports using these assays cannot be compared because interassay test correlation has not been documented. The variability of HPV-16 DNA quantitation was assessed by comparing three real-time PCR assays (HPV-16 L1, HPV-16 E6, and HPV-16 E6 PG) applied on 144 genital samples (125 cervicovaginal lavages and 19 specimens collected using vaginal tampons) obtained from 84 women (66 HIV seropositive and 18 HIV seronegative). Correlation was greater between the HPV-16 E6 assays [correlation coefficient () = 0.92] than between each E6 assay and HPV-16 L1 assay ( = 0.83 and 0.84, respectively). The median HPV-16 copies measured by HPV-16 E6 PG (14,609 HPV-16 copies/2 µL sample) and HPV-16 E6 (18,846 HPV-16 copies/2 µL) were similar (P = 0.27) but were both greater than the median HPV-16 copies measured with the L1 assay (4,124 HPV-16 copies/2 µL; P < 0.001). Correlations between HPV-16 E6 assays were similar for samples containing non-European ( = 0.93) or European ( = 0.95) variants. However, the correlation between HPV-16 L1 and HPV-16 E6 PG or HPV-16 E6 was lower for specimens containing non-European variants ( = 0.80 and 0.76, respectively) compared with specimens containing European variants ( > 0.85). HPV-16 DNA quantity estimated with the three assays was comparable although lower with the HPV-16 L1 assay. The level of correlation depended on viral polymorphism, viral load, and cervical disease status.

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				N. Azizi, J. Brazete, C. Hankins, D. Money, J. Fontaine, A. Koushik, A. Rachlis, K. Pourreaux, A. Ferenczy, E. Franco, et al. Influence of human papillomavirus type 16 (HPV-16) E2 polymorphism on quantification of HPV-16 episomal and integrated DNA in cervicovaginal lavages from women with cervical intraepithelial neoplasia J. Gen. Virol., July 1, 2008; 89(7): 1716 - 1728. [Abstract] [Full Text] [PDF]