建立一种磁微粒化学发光免疫分析法检测肌酸激酶MM型的方法  

Development of a magnetic particle chemiluminescence immunoassay for creatine kinase MM

作　　者：张淑 张春燕[2] 周裕军 许秀丽 程昱璇 舒杨 吴士文[1] 田亚平[2] Zhang Shu;Zhang Chunyan;Zhou Yujun;Xu Xiuli;Cheng Yuxuan;Shu Yang;Wu Shiwen;Tian Yaping(Department of Neurology,First Medical Center of Chinese PLA General Hospital,Beijing 100853,China;Research Center for Birth Defects Prevention and Control Technology,Chinese PLA General Hospital,Beijing 100853,China;Beijing Diagreat Biotechnology Co.,Ltd.,Beijing 101111,China)

机构地区：[1]解放军总医院第一医学中心神经内科医学部,北京100853 [2]解放军总医院医学创新研究部出生缺陷防控技术研究中心,北京100853 [3]北京丹大生物技术有限公司,北京101111

出　　处：《中华检验医学杂志》2024年第6期667-671,共5页Chinese Journal of Laboratory Medicine

基　　金：国家重点研发计划(2022YFC2703600)。

摘　　要：目的自主建立一种快速、高灵敏度,高通量、自动化测定肌酸激酶MM型(CK-MM)同工酶的方法。方法收集2022年1—12月解放军总医院正常新生儿干血斑样本998份,同期就诊的经基因和临床确诊Duchenne型肌营养不良(DMD)患者干血斑样本75份,采用双抗体夹心磁微粒化学发光免疫分析法,测定CK-MM含量。分析该检测方法空白限、交叉反应率、回收率、精密度、钩状效应等性能,并评价其临床符合性。结果该方法空白检测限和检测限分别为17.4和39.3 ng/ml,对肌酸肌酶MB型同工酶(CK-MB)和肌酸激酶BB型同工酶(CK-BB)的交叉反应率分别为0.2%和0.02%。批内和批间变异系数均<1%,回收率为100.32%(95.4%~103.5%),在<50000 ng/ml的范围内无钩状效应出现。新生儿干血斑CK-MM为(27.1±1.0)ng/ml,DMD干血斑CK-MM为(3720.0±300.5)ng/ml。DMD干血斑CK-MM和血清样本CK活性相关性良好(r=0.91,P<0.001),临床符合性好。结论成功建立了一种特异性高、稳定性好的定量测定CK-MM的方法,可满足临床需求,具有较好的应用价值。Objectives Aimed to establish a rapid,high-throughput,automated method for determining the creatine kinase(CK-MM)isoform levels.Methods Magnetic beads labeled with anti-CK-MM antibodies were combined with alkaline phosphatase-based chemiluminescence detection.Clinical and diagnostic performance validation of the assay was determined by analysis of 998 and 75 dried blood spot samples from healthy newborns and Duchenne muscular dystrophy(DMD)patients,respectively,and the CK activity was also determined.The blank and detection limits,cross-reactivity,recovery rate of the method,intra-and inter-assay coefficient,and the hook effect were evaluated.Results Blank and detection limits were 17.4 and 39.3 ng/ml,respectively.Cross-reactivity toward CK-MB and CK-BB isoforms was 0.2%and 0.02%,respectively.Intra-and inter-assay coefficients of variation were<1%.Mean recovery was 100.32%,with no hook effect in CK-MM levels<50000 ng/ml.Overall,the mean CK-MM concentrations in newborns and DMD patients were(27.05±0.97)and(3720±300.5)ng/ml,respectively.A significant positive correlation between the dried blood spot detected CK-MM levels and total CK enzyme activity,evaluated in corresponding serum samples from the 75 DMD patients,was observed(r=0.91,P<0.001),which is in good agreement with the clinical.Conclusions An assay for rapid quantitative determination of CK-MM that meets clinical newborn screening requirements was established.It had a good value for application.

关 键 词：肌酸激酶 MM型 肌营养不良 杜氏 新生儿筛查 

分 类 号：R446.6[医药卫生—诊断学]