COBAS Taqman HBV DNA的方法学验证

Evaluation of the COBAS Taqman Hepatitis B Virus DNA Quantitative Test

目的 评估COBAS Taqman HBV DNA分析系统的性能及临床应用。方法依据美国国家临床实验室标准化委员会（NCCLS）制订的临床化学方法评价方案（evaluation protocols），通过精密度、携带率、线性评价、仪器间比对、参考范围确认等试验，对COBAS Taqman HBV DNA分析系统的性能参数进行了评估。结果COBAS Taqman分析系统批内变异6．9％～8．6％，批间变异10．5％～18．7％，总变异10．8％～20．2％，线性范围6．0～10^7IU／ml，回收率98．7％～104．5％。仪器间比对相关性较好（r^2=0．998）。结论COBAS Taqman HBV DNA分析系统性能指标符合要求，操作简便、快捷，动力学范围广，是目前理想的HBV DNA检测方法。通过此次验证，建立了一套简便的核酸检测方法学评价方案，有利于临床实验室标准化操作的实施。

Objectives To evaluate the performance characteristics of the new real-time PCR COBAS Taqman 48 analyzer. Methods According to the evaluation protocols of National Committee for Clinical Laboratory Standards （NCCLS）, COBAS Taqman was assessed by tests of analytical preclsion, carryover, calibration verification and reference range. Resuits The assay showed low intra-assay （6.9%-8.6%） ,interassay （10.5%-18. 7%） and total（10.8%-20. 2）variations, with good linearity and correlation with expected HBV DNA titers over a wide range （6. 0 to 107 IU/ml）. Compared with another analyzer, results among clinical specimens yielded good correlation （r^2= 0. 998）. Conclusion The performance characteristics of the COBAS Taqman HBV DNA test demonstrated that it provides a sensitive and reliable method for quantitation of HBV DNA levels in the routine molecular laboratory. Through this validation,a set of explicit nucleic acid evaluation protocols has been established in favor of the implementation of the clinical laboratory standardization operation.