动物造模,滋病模型 z-bio 2021-01-18 294

　　Objective: To establish droplet digital PCR (ddPCR) as an important indicator for accurately detecting SIVDNA content in AIDS animal model cells and tissues, so as to evaluate the hidden viruses in reservoirs in real time and dynamically.

　　Method: According to the mature qPCR method, optimize the reaction conditions of droplet dPCR to determine the best annealing temperature. The detection range of ddPCR is determined by detecting 10-fold serial dilutions of the pGEM-SIVgag477 plasmid standard. In addition, the stability of the detection method is judged by testing multiple DNA samples multiple times.

　　Result: The annealing temperature of ddPCR is 60°C, and the detection range is 0.3-3.96×104 copies/μL. The droplet dPCR and qPCR have a good correlation, and the correlation coefficient is r = 0.9981.

　　Conclusion: This study established an absolute quantification method for SIV virus DNA based on droplet dPCR. This can effectively perform dynamic absolute quantitative analysis of viral DNA load in cell and tissue virus library samples, thereby significantly improving the evaluation of model applications.