Vidas hiv duo quick

Serological testing performed at inclusion used a Genzyme Virotech ELISA kit in 2011/an Euroimmun ELISA kit in 2012 (due to hospital acquisition policy), as well as an Euroimmun Western blot kit. The ELISA test was repeated three months post-treatment. To exclude other infections that may cause cross-reactivity, the patients were tested for HIV infection (VIDAS^® HIV DUO Quick-BioMérieux SA, Marcy-l’Étoile, France), Mycoplasma pneumoniae infection (Mycoplasma IgM ELISA-Zeus Scientific, Branchburg, NJ, USA), viral hepatitis B (Monolisa™ Hbs Atg ULTRA—Bio-Rad, Marnes-la-Coquette, France), viral hepatitis C (HCV Ab-DIA.PRO, Sesto San Giovanni, Italy), and syphilis (IMMUTREP^®RPR—Omega Diagnostics, Alva, UK). The presence of the rheumatoid factor was also tested by immunoturbidimetric assay using COBAS C 501 Analyzer (Roche Diagnotics, Basel, Swizerland).

A blood sample of 10 mL maximum was collected from each case and sent at room temperature to the IPB within one hour. Blood smears were prepared, stained with 4% Giemsa and analyzed under a light microscope (×100 oil immersion) to detect forms of Plasmodium species. Hemogram using a Horiba ABX Pentra 80 and measure of the C-reactive-protein using an ABX Pentra 400 (Horiba ABX Diagnostics Inc., Montpellier, France) were realized for each patient. In addition, detection of HIV antibodies was performed using the Alere Determine^™ HIV-1/2 Ag/Ab Combo (Orgenics, Israel) and the Vidas HIV Duo Quick (Biomerieux, France), in accordance with the manufacturer's instructions. Routine serologic testing is generally only informative before the age of 18 months if the test result is negative. Then, HIV-1 proviral DNA PCR (Biocentric, Bandol, France) was performed using the ABI PRISM 7000 real-time PCR system (Applied Biosystems, California, USA) in children younger than 18 months with positive HIV serologic test results. In children older than 18 months of age, HIV-1 and/or HIV-2 Western-blots (New Lav Blot I and II, Biorad, USA) were used to confirm seropositive anti-HIV-1 and/or anti-HIV-2 responses.

The VIDAS HIV Duo Quick (bioMérieux) was used as the index test for the qualitative detection of antibodies to HIV. DBS specimens were punched (1 X 6 mm) using a pneumatic Dried Blood Spot Punch System (Analytical Sales and Services Inc.) into 2 mL 96-well polypropylene plates (ThermoFisher Scientific). DBS punches were eluted in 200 µL of DPBS pH 7.4 containing 0.5% BSA and 0.05% Tween 20 at room temperature for 1 h with agitation (1000 rpm). The elution buffer is based on some of our previous work^{9 (link)}. The punching and elution protocol was carried out according to the Avioq HIV-1 Microelisa System package insert and took into consideration the minimal input volume for the VIDAS HIV Duo Quick test. Afterwards, all remaining DBS eluate was transferred into HIV6 Strips (bioMérieux) and processed by an automated VIDAS benchtop immunoanalyzer (bioMérieux). A test value < 0.25 and ≥ 0.25 was interpreted as negative and positive respectively unless stated otherwise. Test values (TV) were calculated as follows: TV = relative fluorescent value/background fluorescence value.