Nuclisens hiv 1 rna qt assay

We analyzed a cohort of 21 HIC subjects followed-up at the Instituto Nacional de Infectologia Evandro Chagas (INI) in Rio de Janeiro, Brazil. All HIC maintained RNA VL of < 2000 copies/ml without antiretroviral therapy for at least 5 years and were subdivided in two sub-groups: EC (n = 13) when most (≥ 70%) plasma VL determinations were below the limit of detection (LOD), and VC (n = 8) when most (≥ 70%) VL determinations were > LOD and < 2000 copies/ml. The limit of detection of plasma VL determinations varied over the follow-up period in according to the Brazilian Ministry of Health guidelines, with methodologies being updated overtime to improve sensitivity: Nuclisens HIV-1 RNA QT assay (Organon Teknika, Durham, NC, limit of detection: 80 copies/mL) from 1999 to 2007; the Versant HIV-1 3.0 RNA assay (bDNA 3.0, Siemens, Tarrytown, NY, limit of detection: 50 copies/mL) from 2007 to 2013; and the Abbott RealTime HIV-1 assay (Abbott Laboratories, Wiesbaden, Germany, limit of detection: 40 copies/mL) from 2013 to until today. Virological and immunological characteristics of these subjects were described in detail in previous studies [72 , 73 (link)]. Two groups of ART-suppressed subjects (ART, n = 8) and healthy HIV-1-uninfected subjects (NEG, n = 10) were used as controls.

Absolute CD4⁺ T cell counts were obtained using the MultiTest TruCount-kit and MultiSet software with a FACSCalibur flow cytometer (BD Biosciences, California, USA). Plasma VL was measured using the Nuclisens HIV-1 RNA QT assay (Organon Teknika, North Carolina, USA; limit of detection: 80 copies/ml) from 1999 to 2008, the Versant HIV-1 3.0 RNA assay (bDNA 3.0, Siemens, New York, USA; limit of detection: 50 copies/mL) from 2008 to 2013, and the Abbott RealTime HIV-1 assay (Abbott Laboratories, Wiesbaden, Germany; limit of detection: 40 copies/mL) from 2013 to 2016.

Absolute CD4+ T cell counts were obtained using the MultiTest TruCount-kit and the MultiSet software on a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA). Plasma VL was measured according to the Brazilian Ministry of Health guidelines, with methodologies being updated over time to improve sensitivity: Nuclisens HIV-1 RNA QT assay (Organon Teknika, Durham, NC, USA, limit of detection: 80 copies/mL) from 1999 to 2007; the Versant HIV-1 3.0 RNA assay (bDNA 3.0, Siemens, Tarrytown, NY, USA, limit of detection: 50 copies/mL) from 2007 to 2013; and the Abbott RealTime HIV-1 assay (Abbott Laboratories, Wiesbaden, Germany, limit of detection: 40 copies/mL) from 2013 until now.