HIV-1 subtyping using gag/env heteroduplex mobility assay and peptide enzyme-linked immunosorbent assay.
Two HIV-1 subtypes have accounted for virtually all infections in Thailand: subtype B', found mainly in injection drug users (IDUs), and CRF01_AE (initially subtype E), found in over 90% of sexually infected persons and increasingly in IDUs in recent years. During 1997-1998, 227 blood samples were collected from HIV-1 infected individuals consisting of 92 mothers, 35 children and 100 IDUs. The blood samples were subtyped by heteroduplex mobility assay (HMA) and peptide enzyme-linked immunosorbent assay (PEIA). Using gag and env HMA, CRF01_AE and subtype B' accounted for 96-97% and 3-4% of both the mothers and the children, respectively. In the IDU group, 10% of the plasma samples could only be performed by gag HMA and gave the result as CRF01_AE. CRF01_AE and subtype B' using PEIA accounted for 67% and 33% of the IDUs. There was 100% concordance of the results between gag HMA and env HMA. Ninety-five percentages of concordant results were observed between HMA and PEIA. Of the 6/134 (5%) subjects with discordant results, nucleotide sequencing, used as a gold standard, confirmed the HMA result. In this study, HIV-1 was successfully genotyped by HMA and PEIA. However, a comparison of the subtyping results between HMA and PEIA revealed that HMA was slightly more accurate than PEIA.
Simparak
Wantanee
W
Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand.
Kositanont
Uraiwan
U
Sutthent
Ruengpung
R
Wasinrapee
Punneeporn
P
Chaowanachan
Thanyanan
T
Wasi
Chantapong
C
D004279:DNA, Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D024202:Electrophoretic Mobility Shift Assay; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D015752:Genes, env / Q000235:genetics / Q000276:immunology*; D015750:Genes, gag / Q000235:genetics / Q000276:immunology*; D015658:HIV Infections / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000145:classification* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D020180:Heteroduplex Analysis / Q000379:methods; D006801:Humans; D016130:Immunophenotyping; D007223:Infant; D008297:Male; D010455:Peptides / Q000276:immunology; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D013785:Thailand / Q000453:epidemiology
D004279:DNA, Viral; D010455:Peptides