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Early detection of IgA specific antibodies in HIV-1 infected children by peptide-ELISA and peptide time-resolved fluoro-immunoassay
Authors:V. Lombardi  M. Caniglia  G. Scarlatti  M. Jansson  A. Plebani  P. D'Argenio  S. Scaccia  H. Wigzell  P. Rossi
Affiliation:(1) Department of Immunology, Karolinska Institute, Box 60 400, S-104 01 Stockholm, Sweden;(2) Department of Paediatrics, II University of Rome "ldquo"Tor Vergata"rdquo", Rome, Italy;(3) Bambino Gesu' Childrens Hospital, Rome, Italy;(4) Department of Virology, Karolinska Institute, Stockholm, Sweden;(5) Department of Paediatrics I, University of Milan, Milan, Italy
Abstract:The presence of specific IgA antibodies in sera from 25 infants born to HIV-1 seropositive mothers was investigated by peptide-ELISA and peptide time-resolved fluoro-immunoassay (TR-FIA). The infants had been monitored at different times after birth for clinical signs and/or symptoms of HIV-1 infection and for detection of HIV-1 in lymphocyte cultures. Serum samples had also been tested for HIV-1 IgG antibodies by commercial ELISA and Western blot and for p24 antigen. Eleven of 25 children were then identified as infected. IgA detection was performed after rProtein G treatment to remove interfering IgG. In the infected group, IgA specific antibodies to a synthetic peptide representing a highly conserved region of the transmembrane glycoprotein gp41 (env: 594–613) were detected in 27 (73%) out of 37 serum samples (9 of 11 children) by the peptide-ELISA test. IgA specific antibodies to the same peptide were found in 30 (81%) sera (9 of 11 children) by the peptide-TR-FIA. Specific HIV-1 IgA antibodies were detected as early as 2 months of age in serum samples from five out of seven children (71% sensitivity) using peptide-ELISA and from six out of seven (86% sensitivity) by peptide-TR-FIA. Conversely, IgA specific antibodies to HIV-1 were absent in two infected children as well as in the sera of all uninfected children tested during the follow up period. Since maternal IgA does not cross the placenta, IgA detection in the serum of the infant is indicative of HIV-1 infection. Indeed, the early demonstration of HIV-1 IgA antibodies in infected infants shows that both peptide-ELISA and peptide-TR-FIA can be used for an early diagnosis of HIV-1 infection.
Keywords:Mother-to-child transmission  HIV-1  IgA antibodies  Peptide-ELISA  Time-resolved fluoro-immunoassay
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