PNA, probe, primer and method for detecting K-ras gene mutation

Abstract

The invention discloses PNA, probe and primer for testing K-ras genetic mutation, and a method for quantitatively testing the K-ras genetic mutation in real time by the PNA, the probe and the primer. The PNA is hybridized with the 12th codon and / or the 13th codon in the first exon of wild K-ras gene; and the PNA is combined with real-time quantitative PCR to quantitatively test the mutation amount of the K-ras gene in real time. The method has the advantages of simple and convenient operation, high sensitivity and high specificity, and can be applied to early auxiliary diagnosis for pancreatic cancer and other diseases.

Description

technical field [0001] The invention relates to the detection of K-ras gene mutation, in particular to a PNA, probe, primer and method for detecting K-ras gene mutation. Background technique [0002] K-ras gene (GenBank: NC_000012) mutation plays an important role in the occurrence and development of pancreatic cancer. It has been reported that point mutations in codon 12, 13 or 61 of the K-ras gene can be detected in pancreatic cancer tissue, pancreatic juice or feces, and blood (Takahashi, et al. Gastrointestinal Endo 2005.61: 76-9; Luigi L, et al. Oncogene 2002.21:4301-6; TadaM, et al. Cancer Res 1993.53:2472-4; Lu XH, et al.Natl Med J China2001.81:1050-3; N van Heek, et al.J.Clin.Pathol. 2005.58: 1315-1320). [0003] Studies have shown that detection of K-ras gene mutation in somatic DNA can be used as a method for early diagnosis of pancreatic cancer. However, in some experiments, it was found that K-ras gene mutations also exist in benign diseases such as chronic pa...

AI Technical Summary

Problems solved by technology

[0005] (1) The steps are cumbersome and it takes 2 days to complete the identification;

[0006] (2) It can only be used for qualitative research, that is, it can only be determined whether there is a K-ras gene mutation. If quantitative research is required, other methods must be used for further testing;

[0007] (3) False positives caused by incomplete first round of enzyme digestion

[0014] (1) Each specific primer or probe can only be identified for a certain type of gene mutation, and there are more than ten types of mutations in the 12th and 13th codons of the K-ras gene, which need to be detected one by one;

[0015] (2) There may be false positives due to single base mismatches

Images