人肌酸激酶同工酶MB(质量法)化学发光免疫分析定量检测方法的建立及性能评价

Establishment and performance evaluation of the quantitative detection for creatine kinase MB(Mass) based on chemiluminescent immunoassay

利用化学发光免疫分析技术,建立一种人肌酸激酶同工酶MB（CK-MB）定量检测方法。采用基于化学发光免疫分析的双抗体夹心法（以纳米磁微粒为固相载体包被抗人CK-MB抗体,以另一抗体交联碱性磷酸酶为标记物）研制CK-MB化学发光免疫定量检测试剂盒。并进行一系列性能评估及临床相关性试验。该检测方法的线性范围为（0.1-300）ng/ml;空白限为0.1 ng/ml;批内精密度和批间精密度均小于5%;准确度：添加回收率在（100±5）%以内;在正常人血清中添加100 ng/ml的CK-BB及10000 ng/ml的CK-MM,对测定结果没有产生影响;当样本中存在高浓度的甘油三酯、血红蛋白、胆红素以及RF和HAMA时测试偏差在±15%范围以内;试剂在37℃放置72h后。稳定性良好,各性能指标均达到要求;相关性试验结果r〉0.98,一致性结果良好。参考值范围为：（0.42-6.34）ng/ml。本研究建立的CK-MB化学发光免疫分析定量检测方法的各项性能指标均达到了临床检测要求,可用于临床血清CK-MB检测。

This study aims to establish a quantitative detection method for CK-MB（Mass）by chemiluminescent immunoassay.We used double ＂sandwich＂ immunoassay method to develop a CK-MB chemilumineseent quantitative detection kit,in which one anti-human CK-MB antibody coated on the paramagnetic particle was used for capturing,and the other labeled with alkaline phosphatase was used for tracing.The analytical performance of the established assay was further evaluated by a series of analyses.The linear range of the CK-MB quantitative detection kit was（0.1-300）ng/ml.The limit of blank was 0.1 ng/ml.The within-run%CV and between-run%CV are all〈5%.The spiking recovery for accuracy was（100±5）%.No cross-reactivity was observed when CK-BB（100 ng/ml）and CK-MM（10000 ng/ml）were added to normal serum.No significant interference was found with high concentration of bilirubin,triglycerides,hemoglobin,HAMA and RF in the sample.The thermal stability result showed that the kit was stable at 37℃ for 72 hours.The correlation coefficient of clinical comparison test with Beckman ACCESS was r〉0.98 and the consistency analysis with the Bland-altman was〉 95%.The reference range is（0.42-6.34）ng/ml.Taken together,the chemiluminescent immunoassay established in this study reaches the satisfaction of a diagnostic kit and could be applied for the detection of human CK-MB in serum.