Research on ligase-ELISA method for detecting K-ras gene mutations
10.3969/j.issn.1671-8348.2018.02.024
- VernacularTitle:连接酶-ELISA检测K-ras基因突变方法研究
- Author:
Na XIAO
1
;
Yitong TANG
;
Haizhong CUI
;
Zhishan LI
;
Jiuming ZOU
Author Information
1. 湖北文理学院医学院
- Keywords:
genes,ras;
mutation;
circulating DNA;
DNA ligases;
enzyme-linked immunosorbent assay
- From:
Chongqing Medicine
2018;47(2):217-219
- CountryChina
- Language:Chinese
-
Abstract:
Objective To research a simple and sensitive K-ras gene mutations detection method in order to be suitable for the routine mutation detection.Methods The corresponding detection locus oligonucleotide probe was designed.By the connection,amplification,labeling and ELISA reaction in probe,the mutation locus genotype was determined by the ELISA reaction detection value.With the six point mutations of G12S,G12R,G12C,G12D,G12A and G12V in 12 codons of K-ras gene as the detection objects,the plasma circulation DNA sample in 72 cases of lung cancer was detected,then the results were compared with those obtained by the direct sequencing.Results Three samples were identified as the G12S,G12R and G12A mutatins by the established method.But no K-ras mutations were detected in the samples by using the direct sequencing,indicating that the direct sequencing had lower sensitivity and was not suitable for the mutation detection of heterogeneous samples such as circulating DNA.Conclusion The simple and sensitive K-ras gene mutation detection method is established and can conduct the routine mutation detection for the heterogeneous samples.
