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Journal of Clinical Microbiology, October 2002, p. 3654-3659, Vol. 40, No. 10
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.10.3654-3659.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Plasma RNA Viral Load in Human Immunodeficiency Virus Type 2 Subtype A and Subtype B Infections

Florence Damond,1* Marie Gueudin,2 Sophie Pueyo,3 Isabelle Farfara,1 David L. Robertson,4 Diane Descamps,1 Geneviève Chène,3 Sophie Matheron,5 Pauline Campa,5 Françoise Brun-Vézinet,1 and François Simon2

Laboratoire de Virologie, INSERM U552,1 Service des Maladies Infectieuses, SMIT A, Hôpital Bichat, 75877 Paris Cedex 18,5 Laboratoire de Virologie, Hôpital Charles Nicolle, 76031 Rouen,2 INSERM U330, Bordeaux, France,3 Zoology Department, Oxford University, Oxford, United Kingdom4

Received 16 January 2002/ Returned for modification 21 March 2002/ Accepted 28 June 2002

Human immunodeficiency virus type 2 (HIV-2) is much less pathogenic than HIV-1, and HIV-2 infection is associated with plasma viral loads significantly lower than those found in HIV-1 infection. We have developed a real-time quantitative PCR method for measuring the HIV-2 RNA load that covers the range of genetic diversity of HIV-2 isolates and that detects extremely low viral loads. Samples from 49 patients were studied. Proviral DNA was first detected and quantified. The strains that were detected were then genotyped: 21 patients were infected with HIV-2 subtype A and 15 patients were infected with HIV-2 subtype B; 1 patient was infected with a highly divergent strain. Env PCR failed for the remaining 12 patients, so subtypes could not be determined. For viral RNA quantification, a stock of HIV-2 strain NIHZ, which was counted by electron microscopy, was used as the standard. Several primer sets targeting the highly conserved gag region were evaluated. Various primer combinations failed to amplify subtype B strains. With the final primer pair selected, which detected both subtype A and subtype B strains, the sensitivity of the assay was 100% at a viral load of 250 copies/ml and 66% at a viral load of 125 copies/ml. We found a correlation between the CD4+-cell count, the clinical stage, and the plasma HIV-2 RNA level. The median plasma HIV-2 RNA value for the 33 asymptomatic patients was 2.14 log10, whereas it was 3.1 log10 for the 16 patients with AIDS (P < 0.01). Proviral DNA was detectable in 18 symptom-free patients with high CD4+-cell counts, in whom viral RNA was undetectable.

* Corresponding author. Mailing address: Laboratoire de Virologie, INSERM U552, Hôpital Bichat-Claude Bernard, 46 rue Henri Huchard, 75877 Paris Cedex 18, France. Phone: 00 33 1 40 25 88 96. Fax: 00 33 1 40 25 67 69. E-mail: florence.damond{at}bch.ap-hop-paris.fr.

Journal of Clinical Microbiology, October 2002, p. 3654-3659, Vol. 40, No. 10
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.10.3654-3659.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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