ASYMMETRIC NUCLEIC ACID SEQUENCE-BASED AMPLIFICATION (ASYMMETRIC-NASBA) : A DIFFERENT METHOD OF CONVENTIONAL NASBA

Nucleic acid sequence-based amplification (NASBA) is a homogeneous, isothermal, in vitro amplification resulting in the amplification of target sequences more than 10(9)-fold within 120 min([1]). The main amplicons of NASBA are RNA, and currently all the detection method are designed for RNA. In this paper, asymmetric NASBA with unequal concentrations of T7 promotor primer and reverse transcription primer was proposed. By optimizing the ratio of two primers and the concentration of dNPTs and NTPs, the amount of single-stranded cDNA in the amplicons from asymmetric NASBA was found increased 12 times more through the real-time detection of molecular beacon specific to cDNA of interest.