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Journal of Clinical Microbiology, February 2000, p. 630-634, Vol. 38, No. 2
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.
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
*
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.
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