Application of High Resolution Melting Method in the Detection of GLP1R Gene Polymorphism Site rs3765467
10.6039/j.issn.1001-0408.2017.17.01
- VernacularTitle:高分辨率熔解曲线法在GLP1R基因多态性位点rs3765467检测中的应用
- Author:
Yuan ZHANG
;
Xia HE
;
Lei ZHONG
;
Junlan CHUAN
;
Enwu LONG
- Keywords:
GLP1R gene;
High resolution melting;
Single nucleotide polymorphism;
Gene detection
- From:
China Pharmacy
2017;28(17):2305-2308
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish the method for the detection of the known glucagon-like peptide 1 receptor (GLP1R) gene mutation site rs3765467(NT_007592.16,position:39065819),and to evaluate its accuracy and practicability. METHODS:Peripheral venous blood samples of 72 healthy subjects were collected in medical examination center of our hospital during Oct. 2015-Feb. 2016. The whole blood DNA was extracted by column extraction method. After amplified by touch down PCR,high reso-lution melting(HRM)method was adopted to analyze amplified product. Sequencing verification by double stranded chain termina-tion method(Sanger sequencing method)was performed for 38 test samples. The results of 2 methods were compared. RESULTS:The results of mutation scanning showed that there were 39065817 and 39065819 polymorphism sites in amplified segments. Four types of mutations were detected by HRM method [GCG/GCG,GCA/GCG or ACG/GCG,GCA/GCA or ACG/ACG,A(G) CA(G)],but 6 types of mutations was detected by Sanger sequencing method [GCG/GCG,ACG/GCG,ACG/ACG,A(G)CA (G),GCA/GCG,GCA/GCA]. CONCLUSIONS:HRM method can identify GCG/GCG and A(G)CA(G)genotype,but can not identify GCA/GCG and ACG/GCG heterozygous mutation,GCA/GCA and ACG/ACG homozygous mutation. The method is not suitable for the detection of single nucleotide polymorphism for multiple neighboring sites. In the detection of single nucleotide mu-tation,economical and simple method should be selected after comprehensive analysis of sequence.
