荧光定量ＲＴ-ＰＣＲ方法,登革热小鼠模型 z-bo 2021-11-29 435

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

　　Methods Firstly, select appropriate specific primers and probes, prepare plasmid standards by molecular biological methods, optimize PCR system and reaction conditions; secondly, evaluate the sensitivity, specificity and stability of the real-time fluorescent quantitative RT-PCR method; finally, The applicability of this method to the serum and tissue samples of mice infected with dengue virus was verified.

　　Results The one-step real-time fluorescence quantitative RT-PCR method was successfully determined. The method is more sensitive and the lowest detection line is 49.6 Copies/μL. The method has good specificity, no non-specific amplification, good stability, and standard deviation of sample Ct value. Both are less than 0.5, and CV is less than 5%. The test results of the dengue virus infection mouse model samples are in line with the experimental expectations.

　　Conclusion　 The QRT-PCR method can be used as a key indicator detection method for the quantitative detection and evaluation of dengue mouse model viruses.