[Cloning the representative strains of China HIV-1 subtypes B, C and E for heteroduplex mobility analysis (HMA)].
OBJECTIVE
To clone the HIV-1 env gene and establish the HMA reagents suitable for use with the Chinese HIV strains.


METHODS
Using Nest-PCR technique, we amplified env gene from representative strains which contain the most proximate consensus of China HIV-1 subtypes B, C and E. The genes were then cloned into pGEM-Teasy vector for HMA subtyping.


RESULTS
The env gene of representative strains of China HIV-1 subtypes B, C and E were amplified and cloned into plasmids. The plasmids were used in HMA and compared with those standard env gene of other countries. The heteroduplex bands obtained with ours env genes were more proximate to homoduplex bands. The homogeneity of the corresponding China HIV-1 subtypes with standard TH14(B), IND868(C), MA959 (C) and TH22(E) are 90.2%(B), 88.3%(C), 83.7(C)% and 92.3%(E) respectively.


CONCLUSIONS
The HMA reagents we established in this study have higher sensitivity in HMA analysis for China HIV-1 strains when compared with the international HMA reagents. The HMA reagents also provide new reference reagents for UNAIDS HIV isolation network.
Xiao
Y
Y
National Center for AIDS Prevention and Control, Beijing 100050.
Yao
J
J
Shao
Y
Y
D000163:Acquired Immunodeficiency Syndrome / Q000453:epidemiology / Q000821:virology*; D002681:China / Q000453:epidemiology; D019976:Cloning, Organism; D015752:Genes, env / Q000235:genetics; D015497:HIV-1 / Q000145:classification* / Q000235:genetics; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction