A new method for detection of single nucleotide polymorphisms with ligase-ELISA

作者：Cui Hai-zhong; Xiao Na; Zhang Yong-ping; Chen Da-gui; Tang Yi-tong; Zhao Xue-hong; Shao Jin-hui

来源：生物学杂志, 2014, (4).

DOI：10.3969/j.issn.2095-1736.2014.04.095

• SNP
• mutation
• genotyping
• ligase
• ELISA

With the discovery of large number of single nucleotide polymorphism ( SNP) related to human diseases and drug therapy , a large number of approaches for parallel genotyping have been developed .However, these methods require either complicated laboratory procedures or expensive instrumentation .None of these procedures is readily performed in clinical laboratories .In this study, we devel-oped a flexible genotyping method that aims to be inexpensive , accurate, and adapted to routine analysis .The process was called ligase reaction with enzyme-linked immunosorbent assay .This assay was successfully applied to the detection of important SNPs of the EGFR gene at loci of c.2573T> G (L858R), c.2582T> A (L861Q), and c.2155 G> T (G719C).This assay exhibited an excellent speci-ficity, with a sensitivity as low as 5%.With agarose gel electrophoresis and ELISA detection , the genotypes were identified by the present of bands and the values of ELISA detection after 18-28 PCR cycles .In conclusion , the developed technology enables accurate identification of sequence variants in a simple and cost-effective manner from heterogeneous samples unambiguously .

• 出版日期2014
• 单位湖北文理学院