诚信声明我声明，所呈交的毕业论文（设计）是本人在老师指导下进行的研究工作及取得的研究成果。据我查证，除了文中特别加以标注和致谢的地方外，论文（设计)中不包含其他人已经发表或撰写过的研究成果，也不包含为获得其他教育机构的学位或证书而使用过的材料。我承诺，论文（设计）中的所有内容均真实、可信。毕业论文（设计）作者（签名）：2023年月日1基于数字液滴PCR技术建立细胞治疗产品中支原体污染的快速检查法【摘要】目的：通过设计数字液滴P℃R引物和探针，优化反应配方、扩增程序等参数：开发针对支原体DNA的数字液滴PCR方法。方法：参照ICHQ2(R1)等指导原则执行方法学验证，考察dP℃R方法的检测限、专属性和耐用性，比较qPCR方法和ddPCR法的定量结果与检测限。结果：建立支原体的ddPCR快速检查法，为外源因子污染快速检测方法在细胞治疗产品中的应用做出示范，对细胞治疗产品的整体质量和安全评价思路和方法提供范例。结论：dPCR法对比目前主流的qPCR法有更低的检测限，弄够检测出更低浓度的支原体DNA。【关键词】口腔支原体；支原体检测；数字液滴PCR技术；qPCR技术2Rapid screening method for mycoplasma contamination incell therapy products based on digital droplet PCR[Abstract]Objective:To develop a digital droplet PCR method for mycoplasmaDNA by designing digital droplet PCR primers and probes and optimising reactionformulations,amplification procedures and other parameters.Methods:Methodological validation was performed with reference to guidelines such as ICHQ2(R1)to examine the detection limits,specificity and durability of ddPCR methods,and to compare the quantitative results and detection limits of gPCR methods andddPCR methods.Results:A rapid ddPCR assay for mycoplasma was established todemonstrate the application of rapid detection methods for exogenous factorcontamination in cell therapy products and to provide an example of ideas andmethods for the overall quality and safety evaluation of cell therapy products.Conclusion:The ddPCR method has a lower detection limit than the currentmainstream gPCR method and is capable of detecting lower concentrations ofmycoplasma DNA.[Keywords]Oral mycoplasma;Mycoplasma detection;Digital droplet PCRtechnique;qPCR technique3