人肌红蛋白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 myoglobin based on chemiluminescent immunoassay

目的 利用化学发光免疫分析技术,建立一种人肌红蛋白（myoglobin,MYO）定量检测方法。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MYO抗体,以另一抗体交联碱性磷酸酶为标记物）研制MYO化学发光免疫定量检测试剂盒。并进行性能评估及临床试验。结果 该检测方法的线性范围为（1.0~2 500）ng/m L;灵敏度为1.0 ng/m L;批内精密度和批间精密度均小于5%;添加回收率在（100±5）%以内（平均回收率101.92%）;当样本中的MYO浓度为40 000 ng/m L时,未发现钩状效应;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,相关性方程为y=0.9837X-8.9256,与Beckman肌红蛋白测定试剂盒（化学发光法）的一致性结果良好。结论 本研究建立的MYO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MYO检测。

Objective To establish a quantitative detection method for myoglobin （MYO） by chemiluminescent immunoassay. Method Applying double ＂sandwich＂ immunoassay method （one anti-human MYO antibody coated on the paramagnetic particle was used for capturing, and the other anti-human MYO antibody labled with alkaline phosphatase was used for tracing） to develop a MYO chemiluminescent quantitative detection kit. The analytical performance of the established assay was further evaluated by a series of analysis. Resluts The linear range of the MYO quantitative detection kit was （1 - 2 500） ng/mL. The limit of blank was 1 ng/mL. The within-run %CV and between-run %CV were all 〈 5%. The spiking recovery for accuracy was 〈 （100 ± 5）%. There was no hook effect when the sample with 40 000 ng/mL MYO. 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%. Conclusion The method of MYO by chemiluminescent immunoassay which established in this study has reached the satisfaction of a diagnostic kit and could be applied for the detection of human MYO in serum.