胆汁螺杆菌,荧光定量PCR z-bio 2021-01-21 288

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

　　 Method: PCR amplification H. bilis conserved gene P17 sequence full length ORF435bp, TA clone, construct plasmid standard pMD-HBP17. Through the quantitative analysis of pMD-HBP17 standards, the reaction system was optimized to detect the sensitivity, specificity and repeatability of the TaqMan probe real-time fluorescent quantitative PCR method. 77 clinical samples were tested by the established qPCR method and compared with conventional PCR. Compare test results.

　　Result: The established qPCR detection method showed good linearity and correlation between the concentration of plasmid DNA of 108-101 copies. 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 77 copies of clinical samples was 14.3%, which was higher than the detection rate of normal PCR (7.8%).

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