动物造模,登革热 z-bio 2021-08-10 489

　　Objective: To evaluate the real-time fluorescent quantitative RT-PCR method of dengue fever virus and apply it to the detection of key indicators of dengue fever mouse model.

　　Method: First, select appropriate specific primers and probes, prepare plasmid standards by molecular biology methods, and optimize the PCR system and reaction conditions. Next, we will evaluate the sensitivity, specificity and stability of the real-time fluorescent quantitative T-PCR method. The applicability of this method to the serum and tissue samples of mice infected with dengue virus was verified.

　　Result: The one-step real-time fluorescent quantitative RT-PCR method was confirmed. This method is more sensitive, and the lowest detection line is 49.6 copies/μL. This method has strong specificity and no non-specific amplification. Stability and sample Ct value are good. All the standard deviations are less than 0.5, and the CV is less than 5? The test results of the dengue virus infection mouse model samples are in line with experimental expectations.

　　Conclusion: QRT-PCR method can be used as an important indicator detection method for the quantitative detection and evaluation of dengue mouse model virus.