Establishment and application of RT-fqPCR method for detection of drug resistance gene in sufu
In this study,three kinds of drug resistance genes(Baca,Emra,Imrp)were screened to design primers according to the abundance of drug resistance genes in the sequencing results of the whole genome of microorganisms in sufu.A non culture method—real-time fluorescent quantitative polymerase chain reaction(RT-fqPCR)was established to detect the possible resistance genes of microorganisms in different sufu samples,the sensitivity,anti-interference ability and repeatability of the method were tested,and finally applied to the detection of drug resistance genes in sufu samples on the market.The results showed that the established RT-fqPCR method had high sensitivity for detecting three drug resistance genes,all reaching 5.4 pg.The addition of soybean genes had no significant impact on the detection results,but had good anti-interference ability.The relative standard devia-tion(RSD)of the repeatability test results were all less than 1.8%,and the repeatability was good.This method was used to detect 47 batches of com-mercial sufu samples,the results showed that the detection rates of Baca,Emra and Imrp in sufu were 96.5%,92.2%and 97.2%,respectively,and the resistance genes of grey sufu,white sufu and red sufu were 84.4%,83.3%and 89.6%,respectively.In conclusion,the established RT-fqPCR method could rapidly detect three kinds of drug resistance genes in different sufu.
万方数据
维普
