Establishment of a method for detecting KRAS gene mutations based on Taqman fluorescent ARMS-PCR technology
Objective To establish a new method based on Taqman's fluorescent ARMS-PCR technique for detecting the KRAS mutations in human paraffin embedding tissue samples,which is a reference value of clinical medication.Methods Specific primers and probes were designed based on the conserved sequences of KRAS and internal reference genes(GAPDH).Taqman's fluorescent ARMS-PCR detection systems and sequencing systems were established for KRAS detection.In this paper,paraffin embedded tissue samples of 200 patients with colorectal cancer(CRC)are detected and compared with gold standard Sanger sequencing.The results of KRAS gene mutation are statistically analyzed,and the consistency of the two detection methods is analyzed.Results This method is able to detect KRAS gene mutations in human paraffin embedding tissue samples.And the results are 100％consistent with Sanger sequencing.Conclusion The method based on Taqman's fluorescent ARMS-PCR established in this research can detect KRAS mutations simply,quickly and accurately,which is valuable for clinical medication and suitable for clinical application.
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