Abstract:

Objective: To establish a fluorescence PCR method using blood specimen for a rapid genetyping and apply it to the single nucleotide polymorphism(SNP) typing of ADRB1 (1165G>C) gene. Methods: The fluorescence PCR method using blood samples for gene amplification was established without extraction of blood genomic DNA, which was different from a traditional way. A total of 1 051 whole-blood samples anticoagulated with ethylene diamine tetraacetic acid (EDTA) were collected for ADRB1 (1165G>C) gene SNP typing. The performance of self-established method was assessed by comparison with results from Sanger sequencing method, and conformity and consistency (Kappa) between the two methods were calculated. Results: The method established in this research had better amplification performance. The overall typing coincidence rate of method established with Sanger sequencing reached 98.98%, and the Kappa consistency coefficient between them was 0.971. Conclusions: Compared with traditional methods, the method established in this research is easy to operate by omitting the step of genomic DNA extraction with low cost and high accuracy. It is suitable for popularization and application to clinical detection relate to genetyping.

Key words: Fluorescent polymerase chain reaction, Direct amplification, ADRB1 gene, Sanger sequencing