Abstract:

Objective To evaluate the precision,accuracy and limit of detection of real-time fluorescence polymerase chain reaction（PCR） for cytochrome P450 2C19（CYP2C19） polymorphisms.Methods DNA samples with different CYP2C19 genotypes（*1/*1,*1/*2,*1/*3 and *1/*17） were determined for 10 times in 1 run for evaluating precision. The results of real-time fluorescence PCR and Sanger sequencing were used and compared for evaluating accuracy. The DNA samples with different genotypes（*1/*1,*1/*2,*1/*3 and *1/*17） were diluted in a 2-fold serial pattern and determined by real-time fluorescence PCR for evaluating limit of detection.Results Real-time fluorescence PCR showed low coefficient of variation（CV） of Ct values（<2.5%）,a low limit of detection of 1.875 ng/μL and accuracy of 100%（20/20） compared to the results of Sanger sequencing,including 6 kinds of CYP2C19 genotypes in 20 cases（1 case of *1/*17,3 cases of *1/*1,5 cases of *1/*2,5 cases of *1/*3,4 cases of *2/*2 and 2 cases of *2/*3）.Conclusions Real-time fluorescence PCR is precise,accurate and suitable for CYP2C19 polymorphisms.

Key words: Cytochrome P450 2C19, Gene polymorphism, Drug-metabolizing enzyme, Real-time fluorescence polymerase chain reaction, Proficiency testing