利用串联质谱技术筛查新生儿溶酶体贮积症的性能验证及应用  

Evaluation the performance of tandem mass spectrometry for newborn screening of multiple lysosomal storage disorders

作　　者：万智慧[1] 刘伟[1] 孔元原[1] 马志军[1] WAN Zhi-hui;LIU Wei;KONG Yuan-yuan;MA Zhi-jun(Department of Clinical Laboratory,Beijing Obstetrics and Gynecology Hospital,Capital Medical University,Beijing,China,100026)

机构地区：[1]首都医科大学附属北京妇产医院检验科

出　　处：《中国优生与遗传杂志》2018年第12期70-72,69,共4页Chinese Journal of Birth Health & Heredity

基　　金：国家自然科学基金-青年科学基金项目(31700881);首都医科大学基础-临床科研合作基金(17JL58)

摘　　要：目的利用串联质谱技术(Tandem Mass Spectrometry,MS/MS)检测6种溶酶体贮积症(lysosomal storage disorders,LSDs):戈谢病(Gaucher)、尼曼匹克病A/B(Niemann-pick-A/B)、克拉伯病(Krabbe)、黏多糖贮积症I型(MPS-I)、法布里病(Fabry)和庞贝病(Pompe)的筛查指标:酸性β-葡萄糖脑苷脂酶(ABG)、酸性鞘磷脂酶(ASM)、β-半乳糖脑苷酯酶(GALC)、a-L-艾杜糖苷酸酶(IDUA)、a-半乳糖苷酶(GLA)和酸性a-葡糖苷酶(GAA)的活性,分析MS/MS检测性能,建立新生儿这6种酶活性的正常参考范围,并通过检测确诊的溶酶体贮积症样本,评估该方法用于新生儿LSDs筛查的可行性。方法 (1)检测溶酶体贮积症筛查试剂盒自带质控品,评估MS/MS检测LSDs的性能;(2)收集1430例未检测的新筛样本进行检测,建立ABG、ASM、GALC、IDUA、GLA和GAA的酶活性的正常参考区间;(3)检测15?例确诊的LSDs样本,评估该方法用于新生儿LSDs筛查的可行性;(4)分析Pompe患儿ERT治疗前后的酶活差异。结果 (1)MS/MS检测6种酶活性的批内和批间精密度分别为:ABG(2.19-3.74%,1.67-2.08%)、ASM(1.97-3.21%,1.80-1.89%)、GALC(4.77-5.87%,3.22-3.35%)、IDUA(2.49-3.0%,0.88-0.92%)、GLA(1.72-1.76%,0.84-1.65%)和GAA(1.15-1.52%,0.64-1.29%),符合精密度要求;(2)初步确定新生儿6种酶活性参考区间分别为ABG(3.12-28.11?μmol/L*h)、ASM(1.57-18.76μmol/L*h)、GALC(0.97-11.23μmol/L*h)、IDUA(2.67-14.12μmol/L*h)、GLA(3.22-21.09μmol/L*h)和GAA(1.81-20.33μmol/L*h);(3)15例LSDs患儿酶活性明显低于参考区间,MS/MS能有效检出Gaucher、Niemann-pick-A/B、Fabry和Pompe患者;(4)接受ERT治疗后,Pompe患儿的GAA活性显著升高。结论 (1)串联质谱筛查方法可用于新生儿LSDs筛查;(2)初步确定了新生儿6种溶酶体酶活性的参考区间;(3)Pompe患儿ERT治疗后GAA活性显著升高,该方法可用于LSDs患儿用药后的酶活监测。Objective:To evaluate the feasibility of tandem mass spectrometry applied in neonatal screening of LSDs(Gaucher,Niemann-pick-A/B,Krabbe,MPS-I,Fabry and Pompe)by detecting the activities of ABG,ASM,GALC,IDUA,GLA and GAA. Methods:(1)Evaluate the performance of tandem mass spectrometry applied in LSDs screening through calculated the inaccuracy.(2)A total of 1430 newborn screening(NBS)samples were detected by tandem mass spectrometry,and were used to establish the normal reference range for the activities of six enzymes according to 0.5 th to 99.5 th percentiles.(3)A total of 15 samples from patients with LSDs were detected for evaluation the feasibility of NBS of LSDs.(4)Analyze the activity of GAA before and after Enzyme Replacement Treatment(ERT)in Pompe patients. Results:(1)The intraassay and interassay precisions were:ABG(2.19-3.74%,1.67-2.08%),ASM(1.97-3.21%,1.80-1.89%),GALC(4.77-5.87%,3.22-3.35%),IDUA(2.49-3.0%,0.88-0.92%),GLA(1.72-1.76%,0.84-1.65%)and GAA(1.15-1.52%,0.64-1.29%),the precision is fine.(2)The normal reference values were ABG(3.12-28.11 μmol/L*h),ASM(1.57-18.76μmol/L*h),GALC(0.97-11.23 μmol/L*h),IDUA(2.67-14.12 μmol/L*h),GLA(3.22-21.09 μmol/L*h)and GAA(1.81-20.33 μmol/L*h);(3)The enzymes of LSD patients were remarkably decreased compared to the normal range;(4)The GAA activity of Pompe patient increased significiantly after ERT. Conclusion:The present study suggest that the tandem mass spectrometry equipment and kits are suitable for implementation and have acceptable performance for NBS of LSDs.

关 键 词：溶酶体贮积症 串联质谱法检测 新生儿疾病筛查 

分 类 号：R722.1[医药卫生—儿科]