Evaluation of HIV-1/HIV-2 immunoblots for detection of HIV-2 antibodies

To evaluate the sensitivity of commercially available HIV-2 immunoblots and to identify the HIV-2 glycoproteins on Western blots.

HIV-2 Western blot (WB) strips commercially available from Diagnostic Biotechnology, Diagnostic Pasteur and Cambridge Biotech and in-house HIV-2 WB strips were investigated by monoclonal HIV-2 gp36 and gp125 antibodies for identification of the glycoproteins. The WB strips and commercially available HIV-1/HIV-2 line immunoassays (LIAs) from Diagnostic Pasteur (PEPTI-LAV 1-2), Diagnostic Biotechnology (version 2.2) and Innogenetics (INNO-LIA HIV-1/HIV-2 ab) were analyzed by seroconversion panels from HIV-2 infected cynomolgus monkeys (Macaca fascicularis) to investigate their sensitivity for detection of HIV-2 antibodies. The LIAs were also investigated by use of 100 HIV-2 antibody positive human sera from Guinea Bissau. The in-house WB strips contained HIV-2/SBL-6669 antigen treated with various concentrations of sodium dodecyl sulphate (SDS, 0-2%) at 37 degrees C or 100 degrees C for various times to obtain gp36 in oligomeric and/or monomeric form.

By use of monoclonal antibodies, WB strips from Diagnostic Biotechnology and Diagnostic Pasteur were shown to contain gp125 as well as monomeric and oligomeric forms of gp36, whereas Cambridge WB strips contained mainly oligomeric gp36 and no detectable gp125. The sensitivity of the WB strips for detection of HIV-2 seroconversion was similar if WB seropositivity was defined as reactivity with p24 and one envelope protein. When the WHO WB criteria were applied requiring reactivity with at least two envelope proteins for positivity, the sensitivity of the WB strips from Diagnostic Biotechnology and Diagnostic Pasteur was retained, whereas the sensitivity of Cambridge Biotech WB strips was reduced. Among 100 HIV-2 antibody positive human sera all were reactive on PEPTI-LAV 1-2 and INNO-LIA HIV-1/HIV-2 ab, but two of the hundred sera failed to react with the HIV-2 synthetic peptide band on Diagnostic Biotechnology version 2.2 WB strips. On in-house WB strips the relation between monomeric and oligomeric gp36 was changed by altering the SDS concentration and the temperature. Thus the monomeric form increased with the SDS concentration and the temperature. The sensitivity for detection of antibodies during seroconversion did not differ between the monomeric and oligomeric forms of gp36.

The sensitivity for detection of HIV-2 antibodies during seroconversion was independent of the oligomeric or monomeric structure of the transmembrane glycoprotein. One of the three commercial WB kits tested had a lower sensitivity for detection of HIV-2 seroconversion compared with the other two kits when the WHO criteria for WB positivity were used.