电喷雾串联质谱法检测干血斑中谷氨酰胺、谷氨酸及其临床初步应用  

Analysis of glutamine and glutamate in dry blood spots with electrospray ionization-tandem mass spectrometry and its clinical application

作　　者：王彦云[1] 程威[1] 洪冬洋 孙云[1] 蒋涛[1] WANG Yanyun;CHENG Wei;HONG Dongyang;SUN Yun;JIANG Tao(Center for Genetic Medicine,Women′s Hospital of Nanjing Medical University,Nanjing Maternity and Child Health Care Hospital,Nanjing 210004,Jiangsu,China)

机构地区：[1]南京医科大学附属妇产医院&南京市妇幼保健院遗传医学中心,南京210004

出　　处：《临床检验杂志》2021年第7期481-484,共4页Chinese Journal of Clinical Laboratory Science

基　　金：国家重点研发计划(2017YFC1001703,2018YFC1002402);南京市卫生局课题(YKK19118)。

摘　　要：目的建立一种采用电喷雾串联质谱法(ESI-MS/MS)检测干血斑样品中谷氨酰胺(Gln)和谷氨酸(Glu)含量的方法,评价其用于筛查新生儿鸟氨酸氨甲酰转移酶缺乏症和氨甲酰磷酸合成酶缺乏症的可行性。方法收集2020年6—9月新生儿筛查足跟血干血斑样品,用非衍生化串联质谱检测试剂盒萃取样品,完成常规串联质谱检测项目后,将残余样品作为Gln和Glu的检测对象,复溶后选择正离子扫描模式进行分析测定,统计分析ESI-MS/MS检测Gln和Glu的正确度、精密度和稳定性,并评价其临床应用。结果Gln在125~1250μmol/L浓度范围内回收率和正确度均良好,回收率为98.01%~107.24%,偏倚为0.56%~4.8%;Glu在250~1250μmol/L浓度范围内回收率良好,回收率为102.06%~113.65%,在250~1250μmol/L浓度范围内正确度良好,偏倚为0.09%~7.67%。Gln精密度良好,Glu精密度稍差。稳定性实验证实在样品复溶前后,Gln差异无统计学意义,但Glu差异有统计学意义。结合Gln和Glu检测结果,6份鸟氨酸氨甲酰转移酶缺乏症确诊患儿干血斑样品检出率为100%,并确定Gln的参考区间为<106.63μmol/L,Glu的参考区间为<188.24μmol/L。结论成功建立了检测Gln和Glu的ESI-MS/MS法,该方法操作简单、成本低,不影响临床常规检测。Objective To establish a electrospray ionization-tandem mass spectrometry(ESI-MS/MS)method for the determination of glutamine(Gln)and glutamate(Glu)in dry blood spots,and provide its feasibility for screening of ornithine transcarbamylase defi-ciency and carbamoyl phosphate synthetase deficiency in neonatals.Methods The dry blood spots samples were collected from new-born during the period from June to September,2020.The dry blood spots samples were processed with Non-derivatized MS/MS kit.Following the routine MS/MS detection,the residual sample was used for the detection of Gln and Glu.The residual samples were dis-solved in extraction solution,then were detected by ESI-MS/MS with positive ion scanning mode.The accuracy,precision,stability of the method were verified and clinical application and evaluated.Results The recovery and accuracy of Gln were good in the range from 125 to 1250μmol/L(recovery:98.01%to 107.24%,bias:0.56%to 4.8%).The recovery of Glu was good in the range from 250 to 1250μmol/L(recovery:102.06%to 113.65%).The accuracy of Gln was good in the rarge from 250 to 1250μmol/L(bias:0.09%~7.67%).The stability test confirmed that no statistically significant difference of Gln levels before and after reconstitution was found,but the difference of Glu levels was statistically significant before and after reconstitution.Combined with the results of Gln and Glu in 6 ornithine transcarbamylase deficiency positive dry blood spots samples,the diagnostic accuracy was 100%.The reference inter-vals of Gln and Glu were set as<106.63μmol/L and<188.24μmol/L respectively.Conclusion The research successfully established a method for detecting Gln and Glu by ESI-MS/MS in DBS,which is simple,low-cost and without interference in routine clinical de-tection.

关 键 词：电喷雾串联质谱 干血斑 谷氨酰胺 谷氨酸 鸟氨酸氨甲酰转移酶缺乏症 氨甲酰磷酸合成酶缺乏症 

分 类 号：R446[医药卫生—诊断学]