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DNA Nucleic Acid Sequence-Based Amplification-Based Genotyping for Polymorphism Analysis

2004; Future Science Ltd; Volume: 37; Issue: 4 Linguagem: Inglês

10.2144/04374dd04

1940-9818

Cecile Berard, Marie-Angélique Cazalis, Philippe Leissner, Bruno Mougin,

CRISPR and Genetic Engineering

Nucleic acid sequence-based amplification (NASBA) is a sensitive isothermal transcription-based amplification method known to be a suitable tool for RNA research. We demonstrate that NASBA technology can be applied to single nucleotide polymorphism (SNP) analysis using human genomic DNA as a template. Combination of DNA NASBA with multiplex hybridization of specific molecular beacons makes it possible to unambiguously discriminate the presence of the SNP of interest. This protocol is easy-to-use, robust, and makes it possible to rapidly detect single nucleotide substitutions in clinical or cell line DNA sequences using a large range of DNA input. Such a real-time genotyping DNA NASBA assay can find broad application in clinical diagnostics.