Highly sensitive plasma RNA quantification by real-time PCR in HIV-2 group A and group B infection
- PMID: 24008204
- DOI: 10.1016/j.jcv.2013.08.003
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.
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.
Keywords: HIV-2; Natural history; Viral load.
Copyright © 2013 Elsevier B.V. All rights reserved.
Similar articles
-
New Highly Sensitive Real-Time PCR Assay for HIV-2 Group A and Group B DNA Quantification.J Clin Microbiol. 2017 Sep;55(9):2850-2857. doi: 10.1128/JCM.00755-17. Epub 2017 Jul 12. J Clin Microbiol. 2017. PMID: 28701422 Free PMC article.
-
Validation and clinical use of a sensitive HIV-2 viral load assay that uses a whole virus internal control.J Clin Virol. 2013 Dec;58 Suppl 1:e127-33. doi: 10.1016/j.jcv.2013.09.007. Epub 2013 Sep 13. J Clin Virol. 2013. PMID: 24342472
-
Quantitation of HIV-1 RNA levels in plasma and CSF of asymptomatic HIV-1 infected patients from South India using a TaqMan real time PCR assay.J Clin Virol. 2007 May;39(1):9-15. doi: 10.1016/j.jcv.2006.12.026. Epub 2007 Mar 23. J Clin Virol. 2007. PMID: 17368087
-
Quantification of viral DNA during HIV-1 infection: A review of relevant clinical uses and laboratory methods.Pathol Biol (Paris). 2015 Feb;63(1):53-9. doi: 10.1016/j.patbio.2014.07.007. Epub 2014 Sep 8. Pathol Biol (Paris). 2015. PMID: 25201144 Review.
-
Branched DNA for quantification of viral load.Immunol Invest. 1997 Jan-Feb;26(1-2):9-13. doi: 10.3109/08820139709048911. Immunol Invest. 1997. PMID: 9037608 Review.
Cited by
-
Discrepant amplification results during the development of an assay leads to reclassification of two AIDS reagent repository HIV-2 isolates as HIV-1.PLoS One. 2014 May 5;9(5):e96554. doi: 10.1371/journal.pone.0096554. eCollection 2014. PLoS One. 2014. PMID: 24797800 Free PMC article.
-
Assessment of the Cavidi ExaVir Load Assay for Monitoring Plasma Viral Load in HIV-2-Infected Patients.J Clin Microbiol. 2017 Aug;55(8):2367-2379. doi: 10.1128/JCM.00235-17. Epub 2017 May 17. J Clin Microbiol. 2017. PMID: 28515216 Free PMC article.
-
A Multiplex PCR Approach for Detecting Dual Infections and Recombinants Involving Major HIV Variants.J Clin Microbiol. 2016 May;54(5):1282-8. doi: 10.1128/JCM.03222-15. Epub 2016 Feb 24. J Clin Microbiol. 2016. PMID: 26912747 Free PMC article.
-
[HIV-2 infection in Senegal: virological failures and resistance to antiretroviral drugs (ARVs)].Pan Afr Med J. 2019 Jul 18;33:222. doi: 10.11604/pamj.2019.33.222.15771. eCollection 2019. Pan Afr Med J. 2019. PMID: 31692792 Free PMC article. French.
-
The first case of HIV-2 in Scotland.Access Microbiol. 2019 Dec 10;2(3):acmi000087. doi: 10.1099/acmi.0.000087. eCollection 2020. Access Microbiol. 2019. PMID: 32974567 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical