微滴式数字PCR诊断检测应用方案.pdf

Emerging Applications of Droplet Digital PCR in In-vitro diagnostics 杨平 | Ping Yang Ph.D. | Associate Product Manager 1 Bio-Rad基因检测完整解决方案 PCR Droplet Digital PCR, ddPCR 计划申报2类注册证 Real-Time PCR QX200即将获得3类医疗器械注册证 CFX Connect CFX96 Deep Well Dx AutoDG 国械注进20163404214 国械注进20193220316 2 New Solution of Bio-Rad in In-vitro diagnostics Introduction of Digital PCR Droplet Digital PCR Workflow Critical Benefits of ddPCR Emerging Applications of Droplet Digital PCR in In-vitro diagnostics 3 核酸定量技术的革新者 1st Generation 2nd Generation 3rd Generation Gel Electrophoresis Real-Time PCR Droplet Digital PCR (定性/半定量) (相对定量/绝对定量) (绝对定量)  真正的绝对定量，无需外参标准品及标准曲线，数据重复性更好  更高的准确性，不再依赖PCR效率，抑制剂耐受度高，模板适用度广  更高的分辨率，可识别1.1倍浓度差异，适合CNV等类型实验  更高的灵敏度，适合低频突变检测、单细胞基因表达及病原微生物等  一定程度上解决探针法非特异性问题以及染料法引物二聚体问题 4 Bert Vogelstein-最早的数字PCR 5 最早的数字PCR Schematic of Dig-PCR Dig-PCR of DNA from a stool sample Mutant Allele Wild Type Negative Control Positive Control No PCR Product 6 数字PCR时代来临 Digital PCR hits its stride • A sample is diluted and partitioned into hundreds or even millions of separate reaction ch