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Journal of Clinical Microbiology, December 2006, p. 4357-4362, Vol. 44, No. 12
0095-1137/06/$08.00+0     doi:10.1128/JCM.01481-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Quantification of Genital Human Immunodeficiency Virus Type 1 (HIV-1) DNA in Specimens from Women with Low Plasma HIV-1 RNA Levels Typical of HIV-1 Nontransmitters

Departments of Microbiology,¹ Medicine,² Epidemiology,³ Biostatistics, University of Washington, Seattle, Washington; Divisions of,⁴ Human Biology,⁵ Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington,⁶ Department of Medical Microbiology, University of Nairobi, Nairobi, Kenya,⁷ Coast Provincial General Hospital, Mombasa, Kenya⁸

Received 18 July 2006/ Returned for modification 18 September 2006/ Accepted 4 October 2006

Studies of human immunodeficiency virus type 1 (HIV-1) transmission suggest that genital HIV-1 RNA and DNA may both be determinants of HIV-1 infectivity. Despite its potential role in HIV-1 transmission, there are limited quantitative data on genital HIV-1 DNA. Here we validated an in-house real-time PCR method for quantification of HIV-1 DNA in genital specimens. In reactions with 100 genomes to 1 genome isolated from a cell line containing one HIV-1 provirus/cell, this real-time PCR assay is linear and agrees closely with a commercially available real-time PCR assay specific for a cellular housekeeping gene. In mock genital samples spiked with low numbers of HIV-1-infected cells such that the expected HIV-1 DNA copy number/reaction was 100, 10, or 5, the average copy number/reaction was 80.2 (standard deviation [SD], 28.3), 9.1 (SD, 5.4), or 3.1 (SD, 2.1), respectively. We used this method to examine genital HIV-1 DNA levels in specimens from women whose low plasma HIV-1 RNA levels are typical of HIV-1 nontransmitters. The median HIV-1 DNA copy number in endocervical secretions from these women (1.8 HIV-1 DNA copies/10,000 cells) was lower than that for women with higher plasma HIV-1 RNA levels (16.6 HIV-1 DNA copies/10,000 cells) (P = 0.04), as was the median HIV-1 DNA copy number in vaginal secretions (undetectable versus 1.0 HIV-1 DNA copies/10,000 cells). These data suggest that women with low plasma HIV-1 RNA and thus a predicted low risk of HIV-1 transmission have low levels of genital HIV-1 cell-associated virus. The assay described here can be utilized in future efforts to examine the role of cell-associated HIV-1 in transmission.

Published ahead of print on 18 October 2006.

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