Qualitative and quantitative detection of HIV-1 RNA by nucleic acid sequence-based amplification.
OBJECTIVE
To develop a method to detect HIV-1 viral RNA by amplifying a specific nucleic acid sequence.


METHODS
The nucleic acid sequence-based amplification (NASBA) method uses the simultaneous activity of avian myeloblastosis virus reverse transcriptase, T7 RNA polymerase and RNase H to amplify a specific nucleic acid target sequence.


RESULTS
An in vitro cultured HIV-1 stock solution was used to validate the NASBA method and determine the variation in RNA measurement.


CONCLUSIONS
Although NASBA is theoretically capable of specific amplification of RNA or DNA, it is most suitable for amplification of RNA, and therefore for detection of HIV-1 viral RNA.
van Gemen
B
B
Organon Teknika, Boxtel, The Netherlands.
Kievits
T
T
Nara
P
P
Huisman
H G
HG
Jurriaans
S
S
Goudsmit
J
J
Lens
P
P
D005784:Gene Amplification; D015658:HIV Infections / Q000382:microbiology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000706:statistics & numerical data; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014766:Viremia / Q000382:microbiology; D014773:Virology / Q000379:methods* / Q000706:statistics & numerical data
D012367:RNA, Viral