时间分辨荧光免疫分析法检测心肌肌钙蛋白Ⅰ方法学的建立及临床应用

摘要

To establish a method to detect cardiac troponin I by using time-resolved fluoroimmunoassay ( TRFIA) and apply to the clinic.Methods:The assay were measured by TRIFA and double antibody sandwich method .Standard protocols were evaluated with the standard curve , the limit of detection , stability, precision and cross reaction .Healthy reference populations and clinical serum specimens were measured to established the reference interval and evaluated the perspective of the clinical application . Results:The standard curve was Y=7485 .878+1400.924 X with a correlation coefficient of 0.999.The limit of detection was 0.052 ng/ml.The intra-and inter-assay coefficients of variation ( CV) were all <10%.Reference values was <0.14 μg/L.The AUC of ROC curve was 0.971 while the sensitivity was 96.45%,the specificity was 91.43% and the accuracy was 95.69%, with 98.45% of positive predictive value and 82.05%of negative predictive value.The correlation coefficient was 0.993 between our proposed method and the commercially available CLIA kits.There was no significant difference in statistics compared with ECG , CK-MB and cTnT ( P>0.05 ).There was significant difference in statistics compared before and after treatment with AMI ( P<0.001 ) .Conclusion: The TRFIA method for detecting cTnI achieves clinical application standards and may be used for the diagnosis and serosurveillance of acute myocardial infarction patients.%目的：建立时间分辨荧光免疫分析检测心肌肌钙蛋白Ⅰ的方法并应用于临床。方法：采用时间分辨荧光免疫分析及双抗体夹心法，对检测体系的标准曲线、最低检测量、稳定性、精密度和交叉反应等相关性能指标进行评价，通过检测健康体检人员及临床急性心肌梗死患者血清标本并与目前应用于临床的微粒子发光法进行比对，评价TRFIA方法的临床应用可行性。结果：该方法检测肌钙蛋白I的标准曲线Y＝7485.878＋1400.924 X，相关系数为0.999，最低检测限为0.052 ng/ml。批内、批间精密度均＜10％。该方法的正常参考范围为＜0.14μg/L，ROC曲线的AUC为0.971，敏感度96.45％，特异性91.43％，准确度95.69％，阳性预示值98.45％，阴性预示值82.05％。该方法与微粒子发光法具有很强的相关性（ r＝0.993），与ECG、cTnT、CK-MB没有明显的统计学差异（ P＞0.05）。急性心肌梗死患者治疗前后的检测结果具有明显的统计学差异（ P＜0.001）。结论：时间分辨荧光免疫分析法检测肌钙蛋白Ⅰ符合临床诊断的相关要求，可应用于临床急性心肌梗死患者的诊断及疾病检测。