Development of a one-tube multiplex reverse transcriptase-polymerase chain reaction assay for the simultaneous amplification of HIV type 1 group M gag and env heteroduplex mobility assay fragments.
The emergence of intersubtype recombinant HIV-1 isolates has made it imperative to analyze different regions of HIV-1 genomes. For this purpose a one-tube multiplex RT-PCR, coamplifying first-round amplicons that allow amplification of gag and env heteroduplex mobility assay (HMA) fragments from different HIV-1 group M isolates, was developed, starting with plasma samples. The multiplex RT-PCR assay is sensitive: 115 of 136 (84.5%) samples were positive for both gag and env, positive amplification of the gag fragment was observed in 130 of 136 (95.6%) samples, while for the env fragment 119 of 136 (87.5%) tested positive. The multiplex RT-PCR in combination with gag and env HMA makes large-scale HIV-1 subtyping fast, simple, and more economical.
Cham
F
F
Department of Microbiology, Institute of Tropical Medicine, B-2000 Antwerp, Belgium.
Heyndrickx
L
L
Janssens
W
W
Vereecken
K
K
De Houwer
K
K
Coppens
S
S
Van der Auwera
G
G
Whittle
H
H
van der Groen
G
G
D004279:DNA, Viral / Q000097:blood; D015686:Gene Products, env / Q000235:genetics; D015683:Gene Products, gag / Q000235:genetics; D015658:HIV Infections / Q000097:blood / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification; D020180:Heteroduplex Analysis; D006801:Humans; D009692:Nucleic Acid Heteroduplexes; D020133:Reverse Transcriptase Polymerase Chain Reaction
D004279:DNA, Viral; D015686:Gene Products, env; D015683:Gene Products, gag; D009692:Nucleic Acid Heteroduplexes