Isoelectricfocusing and reverse blotting as a diagnostic tool in pediatric HIV infection.

Abstract

Early diagnosis of perinatally acquired HIV-infection is based on either direct HIV detection--by means of viral culture and/or PCR--or anti-HIV antibody detection. However, due to the passive, transplacental passage of maternal immunoglobulin G, antibody detection is nor reliable until 15-18 months of age. In this regard, clonotypic analysis of specific antibodies performed by isoelectricfocusing and reverse blotting (IEF-RB) can be very helpful, as it recognizes possibly different patterns between mother and infant. We used IEF-RB in order to analyze the kinetics of development of anti-HIV antibodies in infants born to seropositive mothers. Sera from ten mother/infant pairs (all mothers were HIV-infected) were retrospectively analyzed in order to detect different patterns, between mother and infant, in anti-gp120 V3-loop clonotype. We diagnosed the real HIV status of the examined infants no later than month 6 and in one case as early as month 2. Considering the small size of sample number, these data are preliminary and should be confirmed by larger scale studies. However, they show IEF-RB, when applied to infants born to seropositive mothers, may be useful in evaluating the infants' dynamics of anti-HIV humoral immune response.