1.Application of heteroduplex mobility assay in genetic subtyping on human immunodeficiency virus type 1.
Liang CHEN ; Yansheng YAN ; Yuwei WENG ; Huirong WANG ; Shouli WU ; Ge CHEN ; Zhaoshuang ZHENG ; Jian ZHENG ; Pingping YAN
Chinese Journal of Epidemiology 2002;23(2):127-130
OBJECTIVEUsing heteroduplex mobility assay (HMA) to subtype human immunodeficiency virus type 1 (HIV-1) for the purpose of understanding HIV-1 subtype epidemic in Fujian province.
METHODSDNA fragments of HIV-1 env gene were amplified from peripheral blood mononuclear cell (PBMC) cocultures of HIV-1 infected individuals by nested polymerase chain reaction (PCR). Heteroduplexs were formed through hybridizing PCR products from the samples and reference plasmid. According to the mobility of heteroduplexs in polyacrylamide gel electrophoresis, HIV-1 subtype from that sample was characterized and further confirmed by nucleotide sequencing analysis.
RESULTSThirteen of 15 (86.67%) samples were successfully subtyped by HMA, except 2 failures. Subtype E and B took up 80% (12/15) and 6.67% (1/15) respectively. Results indicated a high concordance between HMA and nucleotide sequencing analysis and concordance rate was 86.67% (13/15).
CONCLUSIONSSubtype E appeared to be the major epidemic strain of HIV-1 in Fujian. HMA showed the characteristics of fastness, easiness, economic and with high specificity, and can be used in the surviellance for the epidemic strain of HIV-1.
Genotype ; HIV-1 ; classification ; genetics ; Heteroduplex Analysis ; methods ; Polymerase Chain Reaction
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