心肌肌钙蛋白Ⅰ免疫层析试纸条的研制

Development of immunochromatographic strip of human cardiac troponin Ⅰ

目的制备人心肌肌钙蛋白Ⅰ免疫层析试纸条,建立一种快速检测人心肌肌钙蛋白I的检测方法。方法采用柠檬酸三钠还原法制备胶体金,标记肌钙蛋白I单克隆抗体,喷于玻璃纤维纸上制成胶体金结合物垫。将另一株肌钙蛋白I单克隆抗体和抗鼠二抗分别喷于试纸条的检测线和质控线处,组装成试纸条并进行灵敏度、特异性及临床样品测定。结果该试纸条检测灵敏度为1μg/L,15min内可判断结果;与C-反应蛋白、人心肌脂肪酸结合蛋白、肌酸激酶、人心肌肌红蛋白无交叉反应;测定健康人血清样本40例,结果均为阴性;测定14例心肌梗死患者血清,结果均为阳性,且T线显色程度与发光法测定值呈正相关。结论该试纸条灵敏度高、特异性强、操作简便,结果判断直观,可用于急性心肌梗死的早期筛查。

Objective The colloidal gold immunochromatographic test strip was developed in order to establish a quick method for detecting human cardiac troponin I.Methods The colloidal gold was obtained by trisodium citrate reduction method.Colloidal gold particles labelled with monoclonal antibody of cardiac troponin I were used as the detector reagent and coated on some glass fiber and dried.The other monoclonal antibody of cardiac troponin Ⅰ and purified anti-mouse IgG were blotted on the test line and control regions of nitrocellulose membrane,respectively.Then the strip was finished and the detection sensitivity,specificity were measured,and clinical trial was done as well.Results The detection sensitivity of this method was 1 μg/L and the analysis process can be completed within 15 min.There was no cross reaction with C-reactive protein,human heart-type fatty acid-binding protein,human creatine kinase MB and myocardial myoglobin.From 40 blood serum samples of healthy people and 14 of patients with myocardial infarction,the analysis results agreed well with the clinic symptoms.Conclusion This method has high sensitivity and specificity.The procedure of determination is simple and quick without special equipment.It may be used for early diagnosis of AMI.