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Journal of Clinical Microbiology, April 2001, p. 1303-1310, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1303-1310.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Quantification of Human Immunodeficiency Virus Type 1 Proviral Load by a TaqMan Real-Time PCR Assay

Nathalie Désiré,1,* Axelle Dehée,1 Véronique Schneider,1 Christine Jacomet,2 Christophe Goujon,1 Pierre-Marie Girard,2 Willy Rozenbaum,2 and Jean-Claude Nicolas1

Service de Microbiologie1 and Service de Maladies Infectieuses,2 Hôpital Rothschild, 33 Boulevard de Picpus, 75571 Paris Cedex 12, France

Received 20 September 2000/Returned for modification 21 November 2000/Accepted 10 January 2001

Proviral human immunodeficiency virus type 1 (HIV-1) DNA could be a useful marker for exploring viral reservoirs and monitoring antiretroviral treatment, particularly when HIV-1 RNA is undetectable in plasma. A new technique was developed to quantify proviral HIV-1 using a TaqMan real-time PCR assay. One copy of proviral HIV-1 DNA could be detected with 100% sensitivity for five copies and the assay had a range of 6 log10. Reproducibility was evaluated in intra- and interassays using independent extractions of the 8E5 cell line harboring the HIV-1 proviral genome (coefficients of variation [CV], 13 and 27%, respectively) and peripheral blood mononuclear cells (PBMC) from a patient with a mean proviral load of 26 copies per 106 PBMC (CV, 46 and 56%, respectively). The median PBMC proviral load of 21 patients, measured in a cross-sectional study, was determined to be 215 copies per 106 PBMC (range, <10 to 8,381). In a longitudinal study, the proviral load of 15 out of 16 patients with primary infection fell significantly during 1 year of antiretroviral therapy (P = 0.004). In the remaining patient, proviral HIV-1 DNA was detectable but not quantifiable due to a point mutation at the 5' end of the TaqMan probe. No correlation was observed between proviral load and levels of CD4+ cells or HIV-1 RNA in plasma. TaqMan PCR is sensitive and adaptable to a large series of samples. The full interest of monitoring proviral HIV-1 DNA can now be ascertained by its application to the routine monitoring of patients.

* Corresponding author. Mailing address: Service de Microbiologie, Hôpital Rothschild, 33 Boulevard de Picpus, 75571 Paris Cedex 12, France. Phone: 33 (0) 1 40 19 34 33. Fax: 33 (0) 1 40 19 33 35. E-mail: ndesire{at}infobiogen.fr.

Journal of Clinical Microbiology, April 2001, p. 1303-1310, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1303-1310.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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