动物造模 z-bio 2022-03-12 582

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

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

　　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 lowest sensitivity can detect 10 copies/μL, which is 100 times higher than that of ordinary PCR methods; for other common rats There was no non-specific reaction to the virus; the repeatability was good, and the intra-assay and inter-assay coefficients of variation were less than 1%.

　　Conclusion: The TaqMan probe was used to establish a rapid detection of TLV by real-time quantitative PCR method. The method has the characteristics of simple operation, high sensitivity and good specificity.