Highly sensitive plasma RNA quantification by real-time PCR in HIV-2 group A and group B infection

Abstract

Plasma HIV-2 viral load has been reported as predictive of AIDS in HIV-2 infected patient but the lack of sensitivity of the current HIV2 viral load assay is a limitation for the monitoring of the HIV-2-infected patients. ObjectiveTo validate a new quantification assay based on a synthetic HIV-2 RNA transcript and real-time PCR with primers and probes selected in the LTR region, together with high-performance reagents and a protective RNA carrier. Study designWe quantified 23 HIV-2 group A and B supernatants and 58 plasma samples with our TAQMAN-PCR assay and compared the results to those of our previously published in a real time reference PCR performed onto Light Cycler technology, the LC-PCR with a detection of 2.0log10 copies/ml. ResultsThe performance of TAQMAN-PCR was significantly improved, yielding a detection limit of 17 RNA copies/ml. There was a major difference (1–5log10 copies/ml) between LC-PCR and TAQMAN-PCR values for HIV-2 group B supernatants. Twenty-six of 27 plasma samples with levels higher than 2.0log10 copies/ml in LC-PCR were positive by TAQMAN-PCR. Ten of the 31 plasma samples below the LC-PCR detection limit were quantifiable with the TAQMAN-PCR. ConclusionsThe new primers and probe in the LTR region can circumvent HIV-2 diversity, making our method suitable for use in HIV-2 group B-infected patients. Use of a high-performance RT enzyme and RNA carrier protection contributed to improving the detection limit. This study confirms that plasma viral load is lower than 17copies/ml in a large number of HIV-2-infected patients.