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Journal of Clinical Microbiology, February 2000, p. 630-634, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

PCR-Based Assay To Quantify Human Immunodeficiency Virus Type 1 DNA in Peripheral Blood Mononuclear Cells

Cindy Christopherson,1,* Yorda Kidane,1 Brian Conway,2,3 John Krowka,4 Haynes Sheppard,4 and Shirley Kwok1

Roche Molecular Systems, Inc., Alameda, California 945011; University of British Columbia2 and Viridae Clinical Sciences,3 Vancouver, Canada; and Viral and Rickettsial Disease Laboratory, Division of Communicable Disease Control, California Department of Health Services, Berkeley, California 947044

Received 30 July 1999/Returned for modification 20 September 1999/Accepted 1 November 1999

An assay that quantifies the amount of human immunodeficiency virus type 1 (HIV-1) DNA in peripheral blood mononuclear cells has been developed. PCR amplification of the HIV-1 DNA is performed in the presence of an internal quantitation standard, and colorimetric detection of the amplified product is performed with microwell plates. The copies of HIV-1 DNA are normalized to total genomic DNA input. The assay has an analytical sensitivity of 10 input copies per amplification reaction and a three-log detection range. In an analysis of sequential samples from patients on combination therapy, HIV-1 DNA was quantifiable for all individuals tested, including those with undetectable plasma HIV-1 RNA. In a separate study, a comparison of HIV-1 DNA levels was made with a group of long-term survivors and progressors. The mean HIV-1 DNA levels were lower in the long-term survivors than in the progressors (P, 0.04). The mean HIV-1 RNA levels were also lower, but the difference was not statistically significant (P, 0.164). A quantitative DNA assay will provide an additional tool to gain insight into the natural history of infection and the continued efficacy of potent antiretroviral therapies.

* Corresponding author. Mailing address: Department of Infectious Diseases, Roche Molecular Systems, Inc., 1145 Atlantic Ave., Alameda, CA 94501. Phone: (510) 814-2891. Fax: (510) 522-1285. E-mail: Cindy.Christopherson{at}Roche.com.

Journal of Clinical Microbiology, February 2000, p. 630-634, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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