AEGiS-11IAC: Genetic characteristic of HIV-1 in Thailand.

11th International AIDS Conference

Vancouver, British Columbia — July 7-12, 1996

Genetic characteristic of HIV-1 in Thailand.

Int Conf AIDS 1996 Jul 7-12; 11:440 (abstract no. Pub.A.1018)
Auwanit W, Bumisawasdi J, Isarangkura Na Ayuthaya P, Daungchandra S, Saipradit N, Sriwanthana B, Naganawa S, Jayavasu C, Miyamura K, Honda M, Yamazaki M; Department of Medical Science, Ministry of Public Health, Thailand. Fax: 662-5899869. E-mail: watimm@health.moph.go.th.

OBJECTIVE: To characterize field HIV-1 in Thailand genetically and discuss the various factors involved in better understanding the character of HIV to plan and initiate such trials.

METHODS: Blood samples from 352 individuals seropositive for HIV-1 were collected from intra-venous drug users (IVDU), and sexually transmitted disease patients in Thailand. All samples were screened for HIV-1 serotype by V3 domain peptide-base ELISA system. Four peptides represented the principal neutralizing domain (PND) of HIV-1 gene (V3-domain) are PND-E, PND-B, PND-BR and PND-MN. Proviral DNA of 27 blood samples were amplified by using nested PCR that flank the sequences of V3 domain of HIV-1 env region. One of two outer primer pairs, MK603/CO602 or OA3/OD was used for first amplification and inner primer pair EB2/EC2 was used for second PCR. The products were direct sequenced by dye terminator method or were cloned polyclonally and sequenced by dye primer method using automated sequencer. DNA sequences were edited and multiple alignment of amino acid were analyzed by computer software.

RESULTS: By peptide-base ELISA, from 60 samples of IVDUs, 53% were serotype E, 28% were serotype B and 17% were undeternined. From 292 HIV sexual transmission infected samples, 76% were serotype E, 0.7% were serotype B, 8.9% were undetermined and 1% were positive to both PND-E and B.Four undetermined PND-ELISA samples from IVDUs were genotype B by gene sequencing dye terminator method. Three samples demonstrated different HIV-1 subtypes detected by dye terminator and dye primer sequencing methods. From three dual infected samples, the peptide base ELISA results were corresponded to the results of dye terminator method but different from the dye primer method.

CONCLUSION: Two subtypes of HIV-1 (E and B) were presented in Thailand. HIV-1 subtype E commonly found in sexually transmitted group is spreading to IVDU group. Peptide base ELISA can be used for large scale screening of HIV-1 subtype but DNA sequencing by cloning and dye primer sequencing method can demonstrate different suptype of HIV-1 in the same individual.

Keywords: AEGIS, HIV-1, Substance Abuse, Intravenous, Polymerase Chain Reaction, DNA Primers, Enzyme-Linked Immunosorbent Assay, Serotyping, Thailand, Genotype, Peptides, Human, genetics, ICA11

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PubA1018