动物造模,荧光定量检测 z-bio 2021-11-25 812

　　Objective: To establish a clinically rapid and accurate method for detecting suspected Taylor's-like virus of rats (TLV), using TaqMan probe fluorescence quantitative polymerase chain reaction (qPCR) technology, specific Sexual detection of TLV virus nucleic acid.

　　Method: The gene synthesis sequence was used as the template of the plasmid standard. At the same time, the specific sequence was selected at 3622~3729 nt, a pair of primers and TaqMan probes were designed, the reaction system and conditions were optimized, and qPCR amplification was carried out to establish TLV. TaqMan probe qPCR method, and its sensitivity, stability and specificity were evaluated.

　　Results: The established TLV qPCR detection method has a good linear relationship with the standard curve, the R2 value can reach 0.99, and the sensitivity is the lowest. It can detect 10 copies/μL, which is 100 times higher than that of the ordinary PCR method; for other common rats The virus has no non-specific reaction; the repeatability is good, and the coefficient of variation within and between batches is less than 1%.

　　Conclusion: The TaqMan probe is used to establish a fluorescent quantitative PCR method for rapid detection of TLV. This method has the characteristics of simple operation, high sensitivity, and good specificity.