动物造模,荧光定量PCR z-bio 2022-01-27 345

　　Objective: To establish a rapid, sensitive and specific TaqMan probe real-time fluorescent quantitative PCR method for the detection of Helicobacter pylori for the quantitative detection of Helicobacter pylori.

　　Method: H. bilis conservative gene P17 sequence full length ORF435bp PCR amplification, TA cloning, plasmid standard pMD-HBP17 construction. Through the quantitative analysis of pMD-HBP17 standards, the reaction system was optimized to detect the sensitivity, specificity and reproducibility of the TaqMan probe real-time fluorescent quantitative PCR method. 77 clinical samples were tested through the established qPCR method and compared with normal samples. Polymerase chain reaction. Compare the test results.

　　Result: The established qPCR detection method showed good linearity and correlation between 108-101 copies of plasmid DNA concentration. The slope of the obtained standard curve reached -3.46, the correlation coefficient reached 0.999, and the detection sensitivity reached 20. The detection rate of copy 77 clinical samples was 14.3? higher than the detection rate of normal PCR (7.8?.

　　Conclusion: The established H. bilisq PCR detection method has good specificity, high sensitivity and strong stability, and can be used for the quantitative and qualitative detection of Helicobacter pylori.