摘要目的：探讨在ISO15189以及美国病理学家协会（ CAP）实验室认可体系下，建立使用一代测序（Sanger测序法）进行乙型肝炎病毒（HBV）耐药基因检测的项目性能验证标准。方法方法学建立。2012年8至12月收集复旦大学附属华山医院肝炎门诊及住院患者中临床表现HBV耐药患者25例。从测定下限、精密度、正确性、分析特异性、可报告范围/参考范围等方面进行性能评估。测序质量的验证基于整体测序图谱的荧光信号值、性噪比、测序峰图、QV值等评估参数进行。结果可检测出正常背景下10％～20％的突变；有较好的精密度；正确性100％；未发现有明显干扰及交叉污染。结论测序方法的性能验证要结合实际应用。特别是测序分析的质量评估需要针对不同的检测目的以及检测对象建立相关标准并适当进行调整以符合临床需要。本试验方法检测乙型肝炎耐药基因可应用于临床检测。（中华检验医学杂志，2014，37：776-779）

abstractsObjective Under the ISO15189 and America Association of Pathologists ( CAP ) laboratory accreditation system, to establish the performance verification standards for detecting hepatitis B virus resistance gene by sanger sequencing.Methods 25 cases of HBV drug resistance outpatients and inpatients were collected from August 2012 to December in Hepatitis Clinic of Huashan Hospital Affiliated to Fudan University.Analytical performance parameters including analytical sensitivity, precision, accuracy, analytical specificity, reference range/reportable range, etc were evaluated.Sequencing quality evaluation parameters included fluorescence signal intensity overall sequencing chromatogram, signal to noise ratio, trace scores and QV value.Results 10%-20% mutation could be detected under wild-type background. The methool had good precision and accuracy.No obvious interference and cross contamination were observed.Conclusions Performance validation of the sequencing should combine with the practical application.Especially in view of the different detection subjects, and appropriately adjusted to meet the clinical needs.Detection of hepatitis B virus resistance gene by the in the test method in this study can be used in clinical detection.