Nucleic acid sequence-based amplification
Nucleic acid sequence-based amplification (NASBA) is the basis of the NucliSens system of Organon Teknika (now part of bioMerieux) and offers a simple and rapid alternative method for nucleic acid amplification. A description of the technology is available on the web site (http://www.nuclisens.com/). NucliSens represents a synergy of three key technologies – integrating isolation, molecular amplification and detection into an all-in-one system that is targeted at the sensitive and specific determination of nucleic acid sequences. NASBA technology is based on simultaneous enzymatic activity of reverse transcriptase, T7 RNA polymerase, and RNase in combination with two oligonucleotides. It depends on selective primer-template recognition to drive a cyclical, exponential amplification of the target sequence. NASBA has the following
1. Unlike RT-PCR, NASBA is able to selectively amplify RNA sequences in a DNA background, since DNA strands are not melted out. There are no false positive signals due to dead bacteria.
2. It can detect human mRNA sequences without the risk of DNA contamination; no intron flanking primers or Dnase approach is needed. This makes screening for gene expression in oncology or genetic diseases simple and easy.
3. RNA amplification enables direct detection of RNA viruses such as HCV and retroviruses such as HIV.
4. RNA targets permit the detection of live bacterial or viral activity following anti-bacterial/viral therapy.
5. With RNA amplification, it is possible to observe the beginning of cancer cell proliferation through qualitative and quantitative determination of gene activity.
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