Application of High Resolution Melting Method in the Detection of GLP1R Gene Polymorphism Site rs3765467 / 中国药房
China Pharmacy
; (12): 2305-2308, 2017.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-619119
Responsible library:
WPRO
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,position39065819),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.
Full text:
Available
Database:
WPRIM (Western Pacific)
Type of study:
Diagnostic study
Language:
Chinese
Journal:
China Pharmacy
Year:
2017
Document type:
Article