“MS/MS琥珀酰丙酮和非衍生化多种氨基酸、肉碱测定试剂盒”性能验证及应用

An Evaluation the Performance of MS/MS Kit for Newborn Screening

目的对国产“MS/MS琥珀酰丙酮和非衍生化多种氨基酸、肉碱测定试剂盒”(广州市丰华生物工程有限公司,以下简称MS/MS试剂盒)测定新生儿干血滤纸片(dried blood spots specimens,DBS)中氨基酸和肉碱这两类指标的性能进行验证,评估其是否适用于新生儿遗传代谢病筛查。方法(1)通过测定室内质控品和室间质控品,评估精密度和准确度;(2)收集1513例干血滤纸片样本,同时使用MS/MS试剂盒和NeoBase^(TM) Non-derivatized MSMS kit(Perkin Elmer公司)进行检测,分析两种检测试剂盒的一致性,并比较MS/MS试剂盒检测结果和临床诊断结果的一致性;(3)对MS/MS试剂盒检测结果进行分析,初步确定筛查指标的参考范围。结果(1)MS/MS试剂盒检测10种氨基酸的变异系数分别为2.36%~6.85%(CV_(批内))和2.52%~7.44%(CV_(批间)),1种琥珀酰丙酮和15种肉碱的变异系数分别为2.42%~7.12%(CV_(批内))和4.23%~9.54%(CV_(批间))。准确度符合要求;(2)MS/MS试剂盒与NeoBase^(TM) Non-derivatized MSMS kit的一致性评价显示,阳性一致性82.7%、阴性一致性99.5%和总一致性98.7%,kappa=0.854,MS/MS试剂盒检测多种遗传代谢病的灵敏度为100.0%,特异性为96.8%;(3)根据检测结果建立了各筛查指标的初始参考范围,后续随着检测数据的不断积累,将进一步修正切值。结论(1)MS/MS试剂盒检测结果符合实验室性能要求,可应用于新生儿多种遗传代谢病的筛查。(2)MS/MS试剂盒应用于新生儿多种遗传代谢病的筛查需要根据统计结果重新设定参考区间。

Objective To evaluate the feasibility of MS/MS kit applied in newborn screening(NBS)by detecting levels of amino acids and carnitine in blood.Methods(1)Evaluate the performance of MS/MS kit applied in neonatal screening through calculating the inaccuracy and precision of MS/MS kit in detecting levels of amino acids and carnitine of dried blood spots specimens(DBS).(2)1513 samples were detected with MS/MS kit and NeoBase^(TM) Non-derivatized MS/MS kit separately.We evaluated the consistency between these two kits and clinical diagnosis.(3)Determine the initial cutoff value through percentile statistical analysis.Results(1)The coefficient of variation(CV)for amino acids detecting were 2.36%to 6.85%(within-run CV)and 2.52%to 7.44%(between-run CV),respectively.The CV for carnitine detecting were 2.42%to 7.12%(within-run CV)and 4.23%to 9.54%(between-run CV),respectively.The achieved accuracies met requirements.(2)The positive,negative and total consistency of MS/MS kit and NeoBase^(TM) Non-derivatized MSMS kit were 82.7%,99.5%and 98.7%,respectively,with Kappa=0.854.The sensitivity and specificity of MS/MS kit were 100%and 96.8%,respectively.(3)The initial reference range of each screening index was established according to the test results,and the cut-off value would be further modified with the continuous accumulation of test data.Conclusion The present study suggests that the MS/MS kit can be used for NBS of inherited metabolic disease.