UNLABELLED: 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. OBJECTIVE: To 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 DESIGN: We 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 2.0 log10 copies/ml. RESULTS: The performance of TAQMAN-PCR was significantly improved, yielding a detection limit of 17 RNA copies/ml. There was a major difference (1-5 log10 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 2.0 log10 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. CONCLUSIONS: The 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 17 copies/ml in a large number of HIV-2-infected patients.
UNLABELLED: Plasma HIV-2 viral load has been reported as predictive of AIDS in HIV-2 infectedpatient but the lack of sensitivity of the current HIV2 viral load assay is a limitation for the monitoring of the HIV-2-infectedpatients. OBJECTIVE: To 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 DESIGN: We 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 2.0 log10 copies/ml. RESULTS: The performance of TAQMAN-PCR was significantly improved, yielding a detection limit of 17 RNA copies/ml. There was a major difference (1-5 log10 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 2.0 log10 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. CONCLUSIONS: The 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 17 copies/ml in a large number of HIV-2-infectedpatients.
Authors: Didier K Ekouévi; Véronique Avettand-Fènoël; Boris K Tchounga; Patrick A Coffie; Adrien Sawadogo; Daouda Minta; Albert Minga; Serge P Eholie; Jean-Christophe Plantier; Florence Damond; François Dabis; Christine Rouzioux Journal: PLoS One Date: 2015-06-25 Impact factor: 3.240
Authors: Linda L Jagodzinski; Ying Liu; Holly R Hack; Catherine Kibirige; Sheila A Peel; Mark M Manak Journal: PLoS One Date: 2014-05-05 Impact factor: 3.240
Authors: Selly Ba; Ndeye Mery Dia-Badiane; Stephen Edward Hawes; Louise Fortes Deguenonvo; Fatima Sall; Cheikh Tidiane Ndour; Khadim Faye; Fatou Traoré; Macoumba Touré; Marie Pierre Sy; Dana Noelle Raugi; Nancy Berenice Kiviat; Robert Alexander Smith; Moussa Seydi; Papa Salif Sow; Geoffrey Scott Gottlieb Journal: Pan Afr Med J Date: 2019-07-18