AEGiS-14IAC: Comparison between anti-V3 serology and analysis for restriction fragment lengh polymorphism (RFLP) for identification of the Brazilian variant of subtype B (B-GWGR motif).

14th International AIDS Conference

Barcelona, Spain - July 7-12, 2002

Comparison between anti-V3 serology and analysis for restriction fragment lengh polymorphism (RFLP) for identification of the Brazilian variant of subtype B (B-GWGR motif).

Int Conf AIDS 2002 Jul 7-12; 14:(abstract no.. A10014)

Komninakis SC, Fukumori LM, Montanheiro P, Duarte AJ, Casseb J
School of Medicine at Sao Paulo University, Sao Paulo, Brazil

OBJECTIVE: To evaluate anti-V3 antibodies assays and RFLP for identification and discrimination of the Brazilian variant subtype B (B-GWGR motif).

METHODS: Eighty three samples from HIV-1-infected individuals follow-up in the HC/FMUSP and Institute of Infectious Diseases " Emilio Ribas ", were studied. The V3 loop antibodies detection by EIA-solid phase and EIA- Competition using a 15-Aminoacid synthetic peptides, were compared to RFLP using the enzyme FOK 1 (Amersham/Pharmacia) after PCR nested using primers of the NIH (ED5/ED12 and ED31/ES8) and direct DNA sequencing of the V3 region.

RESULTS: Of the 83 samples by EIA solid phase, 48 reacted B-GPGR, 26 to B-GWGR and 9 were indeterminated profile. The 9 indeterminated samples by EIA solid phase were also analyzed through EIA competition. 2 reacted for the subtype B-GPGR, 4 for the subtype B-GWGR, 1 for both subtypes B-GPGR/GWGR and 2 indeterminated pattern. These samples were analyzed through RFLP, 3 showed B-GWGR, one B-GPGR, 1 indeterminated profile and 4 samples had negative PCR. The 83 samples were analysed by RFLP, 36 characterized as subtype B-GPGR, 20 as of the subtype B-GWGR, 18 had indeterminated patterns with non expected bands and 9 samples had negative PCR. We compared the results of the EIA solid phase and of RFLP for the discrimination among the subtype B-GPGR and of the subtype B-GWGR. Thirty-six samples were characterized as subtype B-GPGR by RFLP, compared with 48 by EIA solid phase (75% of concordance). Twenty samples with B-GWGR motif for RFLP, compared with 26 subtyping by EIA solid phase, obtaining an agreement of 77%.

CONCLUSIONS: V3 serology and RFLP present a 75-77% concordance for discrimination of subtype Brazilian variant B-GWGR. Support: FAPESP 97/11502-2 and 00/11868-1

Keywords: AEGIS, Polymorphism, Restriction Fragment Length, HIV-1, Polymorphism (Genetics), Serologic Tests, Variation (Genetics), Polymerase Chain Reaction, AIDS Serodiagnosis, Brazil, immunology, genetics

020707
A10014