Aim: To develop a method to detect HIV-1 viral RNA by amplifying a specific nucleic acid sequence.
Method: 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.
Validation: An in vitro cultured HIV-1 stock solution was used to validate the NASBA method and determine the variation in RNA measurement.
Conclusion: 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.