动物造模,时荧光定量 z-bio 2021-12-23 506

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

　　Method: 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 RT-PCR method. The applicability of this method to serum and tissue samples from mice infected with dengue virus was verified.

　　Result: The step-by-step real-time fluorescence quantitative RT-PCR method was confirmed. This method is more sensitive, and the minimum detection line is 49.6 copies/μL. The method is highly specific and has no non-specific amplification. It has good stability and sample Ct value. All standard deviations are less than 0.5 and CV is less than 5%. The test results of the mouse model samples infected by the dengue virus are in line with the expectations of the experiment.

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