XENONUCLEIC ACID-MEDIATED MULTIPLEX QPCR CLAMPING TECHNOLOGY FOR LUNG CANCER MUTATION DETECTION

摘要

The invention provides a multiplex method for enriching a plurality of target polynucleotide sequences containing genetic mutations associated with lung cancer comprising: (a) providing a biological sample; (b) isolating DNA from said sample; said DNA including said plurality of target polynucleotide sequences containing genetic mutations; (c) providing a plurality of primer probes targeted to said target polynucleotide sequences said primer probes allowing formation of a PCR product; (d) providing a plurality of target specific xenonucleic acid clamps oligomer probes specific for wildtype polynucleotide sequences; so that during the qPCR process only mutant templates are amplified: (e) admixing the plurality of primer probes and the plurality of xenonucleic clamping probes with the target nucleic acid sample; (f) performing a PCR amplification process in reaction solution under hybridization conditions thereby generating multiple amplicons; and (g) detecting said amplicons and wherein said xenonucleic acid clamps have aza-aza, thio-aza and oxy-aza chemical functionality.