Establishment of pyrosequencing method for detection of k-ras mutation

Abstract

Abstract: AIM: To establish a pyrosequencing method for detecting k-ras mutation and to investigate the frequency and characteristic of k-ras of patients with colorectal cancer.METHODS: A total of 10 DNA specimens from the ministry of health clinical test center of China for external quality assessment were prepared for k-ras mutation detecting method verification. A total of 230 DNA specimens from patients with colorectal cancer were used for k-ras mutation detection.RESULTS: The overall positive rate of k-ras gene mutations obtained by pyrosequencing was 14.34% (33/230), and the positive rate of mutations was 10% (23/230) at codon 12 [5.22% (12/230) for 35G>A, 3.04% (7/230) for 35G>T, 1.74% (4/230) for 34G>T] and 4.34% (10/230) at codon 13[0.43% (1/230) for 37G>A, 3.91% (9/230) for 38G>A].CONCLUSION: This pyrosequencing method to detect k-ras mutation is proved to be a rapid, accurate and high throughput method alternative to conventional method, and it can be a preferred option in research and clinical application.

Key words: colorectal cancer, k-ras gene, gene mutation, pyrosequencing

CLC Number:

R969

ZHOU Xia-hui, WU Cheng-jiu, XIAO Le-dong, SUN Li-xian, CAO Zhong. Establishment of pyrosequencing method for detection of k-ras mutation[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2015, 20(4): 430-434.

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