人全外显子组测序IDT捕获探针的性能验证

目的探讨已建立的二代测序技术进行人全外显子组测序项目对变异的检出性能。 方法对4例实验室能力比对验证样本和4例已知结果样本进行全外显子组检测，分析点变异和插入缺失变异，从检测准确度、检测灵敏度、检测特异性、精密度和可报告范围等方面进行性能评估。数据质量控制以目标区域覆盖率、捕获特异性和平均深度为基础。 结果数据输出量大于8G，实现了目标序列区域99.7%以上的覆盖率，捕获特异性86%以上，平均测序深度100×以上，Q30大于90%，对SNV的检出率达100%，50 bp以下Indel检出率达100%。 结论本标准操作在验证范围内对变异的检出能力达到应用的要求。

Performance verification of IDT capture probes for human Whole-exome sequencing

ObjectiveTo investigate the detection performance of Whole-exome sequencing (WES) for calling variants by the established second-generation sequencing technology. MethodsFour CAP proficiency testing samples and 4 known results samples were detected by WES. Point mutations and insertion deletion mutations were analyzed. Detection performance was evaluated in terms of accuracy, sensitivity, specificity, precision and reportable range of calling variants. Data quality control was performed based on coverage of target region, capture specificity and mean depth. ResultsOutput data should not be less than 8G. In this case, coverage of target region is over 99.7%. Capture specificity is over 86%. Mean depth is over 100×. Q30 is over 90%, The detection of single nucleotide variations and small insertion and deletion under 50bp is up to 100%. ConclusionThe standard operating procedure within the scope of validation can meet the application requirement of mutation detection.