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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{triangledown}

Sarah Benki,1,5 R. Scott McClelland,2,3,7 Sandra Emery,5 Jared M. Baeten,2 Barbra A. Richardson,4,6 Ludo Lavreys,2,7 Kishorchandra Mandaliya,8 and Julie Overbaugh5,6*

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.

* Corresponding author. Mailing address: Division of Human Biology, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Mail Stop C3-168, Seattle, WA 98109. Phone: (206) 667-3524. Fax: (206) 667-1535. E-mail: joverbau{at}fhcrc.org.

{triangledown} Published ahead of print on 18 October 2006.

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.



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  • Voronin, Y., Chohan, B., Emerman, M., Overbaugh, J. (2007). Primary Isolates of Human Immunodeficiency Virus Type 1 Are Usually Dominated by the Major Variants Found in Blood. J. Virol. 81: 10232-10241 [Abstract] [Full Text]  


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