Development of an antigen 'sandwich' enzyme immunoassay for the detection of antibodies against HIV-2 by using a biotinylated synthetic peptide of gp36 protein
ENFERMEDADES INFECCIOSAS Y MICROBIOLOGIA CLINICA
Authors: Delahanty-Fernandez, Aurora; Clara Bequer-Ariza, Dunia; Hernandez-Marin, Milenen; Zulueta-Rodriguez, Orlando; Pozo-Pena, Lilliam; Hernandez-Spengler, Idialis; Ramos-Martinez, Grisell; Antonio Valdespino-Diaz, Marcos; Ventura-Paz, Julio
Abstract
Introduction: Among the several existing methods for the detection of antibodies to HIV, the 'sandwich' ELISA is currently the most used. This study aims to assess a biotinylated monomeric synthetic peptide of the glycoprotein trans-membrane gp36 from HIV-2, in a sandwich assay, for the detection of antibodies against this HIV-2 protein. Materials and methods: To perform the assay, plates coated with recombinant protein gp36 at 0.5 mu g/mL and synthetic peptide gp36(5) at 1 mu g/mL were used. The concentration of the biotinylated synthetic peptide (gp36(5)-B) used was 0.1 mu g/mL prepared with a Tris-BSA-NaCI buffer solution and the Streptavidin-Alkaline Phosphatase conjugate diluted 1:30000 prepared with a PBS-Sucrose-BSA solution. Positive serum samples to antibodies against HIV-1 and HIV-2 viruses (88 and 34, respectively) were tested, with 483 negative samples from blood donors and 96 serum samples to assess the analytical specificity. All the samples were tested using the UMELISA HIV 1+2 RECOMBINANT assay, and all positives were confirmed using a DAHIV-BLOT assay. Results: Thirty four samples with antibodies against HIV-2 were assessed as positive for both coating variants. The highest specificity was obtained with the variant using the synthetic peptide gp36(5) in its coating. The antigen 'sandwich' assay developed by using gp36(5)-B enables the detection of antibodies against gp36 protein of HIV-2 (C) 2014 Elsevier Espana, S.L.U. and Sociedad Espanola de Enfermedades Infecciosas y Microbiologia Clinica. All rights reserved.
Frequency of human immunodeficiency virus type-2 in hiv infected patients in Maputo City, Mozambique
VIROLOGY JOURNAL
Authors: Maueia, Cremildo; Costa, Deise; Meggi, Bindiya; Ismael, Nalia; Walle, Carla; Curvo, Raphael; Abreu, Celina; Bhatt, Nilesh; Tanuri, Amilcar; Jani, Ilesh V.; Ferreira, Orlando C., Jr.
Abstract
The HIV/AIDS pandemic is primarily caused by HIV-1. Another virus type, HIV-2, is found mainly in West African countries. We hypothesized that population migration and mobility in Africa may have facilitated the introduction and spreading of HIV-2 in Mozambique. The presence of HIV-2 has important implications for diagnosis and choice of treatment of HIV infection. Hence, the aim of this study was to estimate the prevalence of HIV-2 infection and its genotype in Maputo, Mozambique. HIV-infected individuals (N = 1,200) were consecutively enrolled and screened for IgG antibodies against HIV-1 gp41 and HIV-2 gp36 using peptide-based enzyme immunoassays (pepEIA). Specimens showing reactivity on the HIV-2 pepEIA were further tested using the INNO-LIA immunoblot assay and HIV-2 PCR targeting RT and PR genes. Subtype analysis of HIV-2 was based on the protease gene. After screening with HIV-2 pepEIA 1,168 were non-reactive and 32 were reactive to HIV-2 gp36 peptide. Of this total, 30 specimens were simultaneously reactive to gp41 and gp36 pepEIA while two samples reacted solely to gp36 peptide. Only three specimens containing antibodies against gp36 and gp105 on the INNO-LIA immunoblot assay were found to be positive by PCR to HIV-2 subtype A. The proportion of HIV-2 in Maputo City was 0.25% (90%CI 0.01-0.49). The HIV epidemic in Southern Mozambique is driven by HIV-1, with HIV-2 also circulating at a marginal rate. Surveillance program need to improve HIV-2 diagnosis and consider periodical survey aiming to monitor HIV-2 prevalence in the country.