Vironostika hiv uni form 2 ag ab

HIV serostatus was determined using double ELISA testing (Murex HIV-1.2.O, Abbott Murex, UK and Vironostika^® HIV Uni-form II Ag/Ab, Biomérieux, The Netherlands). CD4+ count was estimated using the FlowCARE™ PLG CD4 (CD45-FITC/CD4-PE) assay (Beckman Coulter, CA, USA) and HIV-1-RNA with RealTime HIV-1 assay (Abbott Laboratories, Abbott Park, Illinois, USA). HCV antibody testing was performed using the Genedia HCV ELISA 3.0 (Green Cross Medical Science, Chungbuk, Korea). HCV RNA was ascertained using the RealTime HCV assay (Abbott Molecular Inc., Des Plaines, IL, USA). Chronic HBV infection was measured by presence of hepatitis B surface antigen (Hepanostika HBsAg Uniform II, Biomérieux, The Netherlands). Plasma glucose, tryglicerides, total cholesterol, HDL cholesterol, LDL cholesterol were measured using an enzymatic methods (Olympus AU400; Olympus Diagnostica, Tokyo, Japan). Plasma insulin was measured by immunoassay (ELISA) using a Bioscience kit (Monobind kit, Monobind Inc., Lake Forest, CA, USA). Homeostasis model assessment (HOMA–IR) was calculated as (fasting plasma glucose (mmol/L) x fasting insulin (μU/mL)/22.5).

Plasma or serum samples from blood collected by venipuncture in each MSM were used for serological testing of HIV infection, as recommended by national guidelines [25 (link)]. HIV-positive status was determined by the national serological testing algorithm using Genscreen ULTRA Combo HIV Ag/Ab test (Bio-Rad Laboratories, Washington, USA), an enzyme immunoassay kit for the simultaneous detection of HIV p24 antigen and antibodies to HV-1 (group M and O) and HIV-2 in serum or plasma. The positive tests were confirmed by an enzyme-linked immunosorbent assay [ELISA] (Vironostika® HIV Uni-Form II Ag/Ab, bioMérieux Marcy l'Etoile, France). Specimens for molecular testing were obtained by inserting a moistened polyester swab into the anal canal, rotating 5 times and then removing. The swab was immediately placed into a sampler tube and frozen at -80°C before the DNA extraction procedure.

Clients’ status was determined by using the reference standard algorithm at the AIDS reference laboratory at ITM, Antwerp, Belgium (Figure 1) on collected plasma samples. All samples were tested by a fourth generation ELISA (Vironostika® HIV Uni-Form II Ag/Ab, bioMérieux, France) and all reactive samples were confirmed by a Line-Immunoassay (LIA, i.e. INNO-LIA™ HIV I/II Score, Innogenetics NV, Ghent, Belgium). Samples with a negative or indeterminate LIA were tested with an antigen-enzyme-immunoassay (Ag-EIA, i.e. INNOTEST HIV Antigen mAb, Innogenetics NV, Ghent, Belgium) to confirm acute infections. In the event that the LIA could not differentiate between HIV-1 and HIV-2, we used an in-house DNA PCR.
Figure 1.

Reference algorithm at the AIDS reference laboratory at the Institute of Tropical Medicine, Antwerp, Belgium.