title: New sensitive one-step real-time duplex PCR method for group A and B HIV-2 RNA load.
abstract: The Agence Nationale de Recherche sur le Sida et les hépatites virales (ANRS) previously developed a widely used method for HIV-1 RNA quantification (Biocentric). Here, we report the development of a new specific and sensitive method for HIV-2 RNA quantification, based on an adaptation of the existing HIV-1 protocol. The new test is based on TaqMan one-step reverse transcription-quantitative PCR (qRT-PCR) targeting two conserved consensus regions of HIV-2 (long terminal repeat [LTR] and gag). Analytic performances were determined in three laboratories. Clinical performances were evaluated on 100 plasma samples from HIV-2-infected patients (groups A, B, and H) by comparison with the assay currently used for the ANRS HIV-2 cohort. The specificity was 100%. Sensitivity was 50 copies/ml (cp/ml) and was optimized to 10 cp/ml. The within-run coefficients of variation in the three laboratories varied from 0.54% to 1.61% at 4 log10 copies/ml and from 7.24% to 14.32% at 2 log10 cp/ml. The between-run coefficients of variation varied from 2.28% to 6.43%. Of the 39 clinical samples below 2 log10 in the current assay, the new test improved the detection or quantification of 17 samples, including eight group B samples. For quantifiable samples, similar loads were obtained with the two assays for group A samples. The median difference between the two assays for group B samples was +0.18 but with greater heterogeneity than for group A. The HIV-2 group H sample had similar results with the two assays. This new assay is highly sensitive and accurately quantifies the most prevalent HIV-2 groups. This test will be useful for monitoring low viral loads in HIV-2-infected patients.
authors.affiliation: Laboratoire de Virologie, AP-HP, Hôpital Necker Enfants Malades, Paris, France EA7327, Université Paris-Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Avettand-Fenoel
authors.affiliation: INSERM, IAME, UMR 1137, Paris, France Université Paris Diderot, Sorbonne Paris Cité, Paris, France AP-HP, Hôpital Bichat-Claude Bernard, Laboratoire de Virologie, Paris, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Damond
authors.affiliation: Laboratoire de Virologie Associé au Centre National de Référence du VIH, Hôpital Charles Nicolle, CHU de Rouen, Rouen, France GRAM, Equipe d'Accueil 2656, Faculté de Médecine-Pharmacie, Institut de Recherche et d'Innovation en Biomédecine, Université de Rouen, Rouen, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Gueudin
authors.affiliation: INSERM, IAME, UMR 1137, Paris, France Université Paris Diderot, Sorbonne Paris Cité, Paris, France AP-HP, Hôpital Bichat-Claude Bernard, Service de Maladies Infectieuses et Tropicales, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Matheron
authors.affiliation: EA7327, Université Paris-Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, France.
authors.forename: Adeline
authors.initials: A
authors.lastname: Mélard
authors.affiliation: INSERM, IAME, UMR 1137, Paris, France Université Paris Diderot, Sorbonne Paris Cité, Paris, France AP-HP, Hôpital Bichat-Claude Bernard, Laboratoire de Virologie, Paris, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Collin
authors.affiliation: INSERM, IAME, UMR 1137, Paris, France Université Paris Diderot, Sorbonne Paris Cité, Paris, France AP-HP, Hôpital Bichat-Claude Bernard, Laboratoire de Virologie, Paris, France.
authors.forename: Diane
authors.initials: D
authors.lastname: Descamps
authors.affiliation: Laboratoire de Virologie, AP-HP, Hôpital Saint-Louis, Paris, France.
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Chaix
authors.affiliation: Laboratoire de Virologie, AP-HP, Hôpital Necker Enfants Malades, Paris, France EA7327, Université Paris-Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Rouzioux
authors.affiliation: Laboratoire de Virologie Associé au Centre National de Référence du VIH, Hôpital Charles Nicolle, CHU de Rouen, Rouen, France GRAM, Equipe d'Accueil 2656, Faculté de Médecine-Pharmacie, Institut de Recherche et d'Innovation en Biomédecine, Université de Rouen, Rouen, France jean-christophe.plantier@univ-rouen.fr.
authors.forename: Jean-Christophe
authors.identifier: http://orcid.org/0000-0001-6417-4083
authors.initials: JC
authors.lastname: Plantier
mesh_terms: D015658:HIV Infections / Q000821:virology*; D015498:HIV-2 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods*