Profile of antigen-specific antibody response detectable by Western blot in relation to diagnostic criteria for human immunodeficiency virus type-1 (HIV-1) infection

Abstract

Background: Variability in the profile of antigen-reactive bands in Western blot for serodiagnosis of human immunodeficiency virus (HIV) infection may result in disagreement regarding interpretation of positive result, due to lack of consensus in the interpretive criteria laid down by various organisations.

Objectives: The objectives of this study were (i) to find out the extent of disagreement over various criteria regarding interpretation of positivity in Western blot and (ii) to review the discordance by retesting the discordant specimens using recombinant antigens as well as by performing repeat Western blot in follow-up specimens.

Study design: A total of 467 specimens from high-risk groups, diagnosed positive for HIV type-1 (HIV-1) infection by the criteria of at least one of the five organisations, viz. Association of State and Public Health Laboratories Directors (ASTPHLD), Consortium for Retrovirus Serology (CRSS), American Red Cross (ARC) and World Health Organisation (WHO), were analysed to find out the extent of discordance between various criteria for interpretation of Western blot positivity. The discordant specimens were subjected to line immunoassay (LIA) using recombinant antigens. Also, follow-up Western blots were performed in case of discordant specimens at 6, 12 and 24 weeks intervals.

Results: We observed that criteria laid down by ASTPHLD, CDC and CRSS scored all the specimens as positive while ARC and WHO criteria scored 13 (2.8%) and 18 (3.8%) of specimens, respectively, as negatives which were detected as positives by other criteria (discordant specimens). The gp41 reactive band was the most frequently missing band, being undetectable in 11.6% of specimens while bands reactive to p24, p31, gp120 and gp160 could not be recorded in 1.9%, 9.4% and 3.2% and 1.5% of specimens, respectively. Testing of the discordant specimens with recombinant antigen preparation and with repeat Western blot in follow-up specimens collected at 6, 12 and 24 weeks demonstrated all bands undetectable in initial Western blot, except 25% of gp41 reactive bands.

Conclusions: It is felt that before selecting any criterion for Western blot positivity, it should be evaluated in the local population at risk for HIV-1 infection with additional or follow-up tests.