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| Establishment and verification of TaqMan-MGB two-color fluorescent probe method for detecting the gene polymorphisms of insulin receptor substrate 1 rs1801278 |
| ZHAO Yangyang1,2 XU Zhihui2 LIU Yan2 WU Rongrong3 LIU Xinguang1 XU Dongping2▲ HAN Jin3▲ |
1.Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics Institute of Biochemistry and Molecular Biology, Guangdong Medical University, Guangdong Province, Dongguan 523808, China;
2.Administration Center of Clinical Research, 302 Military Hospital of China, Beijing 100039, China;
3.Department of Pharmacy, 302 Military Hospital of China, Beijing 100039, China |
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Abstract Objective To establish an experimental method suitable for clinical routine detection of the gene polymorphisms of insulin receptor substrate 1 (IRS1) rs1801278. Methods The peripheral blood samples of 200 healthy controls in 302 Military Hospital of China from March 2015 to June 2016 were collected, the specific primers and fluorescent probe aimed at the polymorphic sites of gene polymorphisms of IRS1 rs1801278 were designed, and the gene polymorphisms of rs1801278 were detected by the TaqMan-MGB real-time fluorescence polymerase chain reaction and validated by gene sequencing method (golden standard). The distribution frequencies of different genotypes of IRS1 rs1801278 in this population were discussed as well. Results The TaqMan-MGB two-color fluorescent probe method could identify 3 genotypes of IRS1 rs1801278, achieving 100% consistence with gene sequencing method. The distribution frequencies of T/T, C/T and C/C genotypes of gene polymorphisms of IRS1 rs1801278 in this population were 0%, 1% and 99%, respectively. Conclusion The established TaqMan-MGB fluorescent probe method based on the specific probe and primer of single nucleotide polymorphisms is rapid and effective in identifying the genotypes of gene polymorphisms of IRS1 rs1801278, the results are reliable, and the operation is easy, but the predictive value for the individualized therapeutic effect of Chinese Han type 2 diabetes patients is limited, which is not recommended for clinical routine practice.
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