CMLC夹心酶联免疫分析的建立及初步应用

Establishment and primary application of sandwich ELISA for human cardiac myosin light chain

目的建立人心肌肌凝蛋白轻链（CMLC）夹心酶联免疫分析方法并初步进行应用研究。方法通过对鼠抗人CMLC单克隆抗体（McAb）的相加指数、亲和常数进行测定和夹心实验,筛选到配对较好的两对CMLC单抗,在此基础上建立了CMLC夹心酶联免疫分析方法,用该方法对心肌梗塞病人和正常人血清CMLC进行测定。结果最佳CMLC单抗包被浓度为5～7μg/mL,CMLC抗原在1～50ng/mL范围内标准曲线线性良好。血清CMLC测定OD值：正常人（20名）为（0.875±0.162）,心肌梗塞病人（30名）为（1.463±0.473）。结论此方法可以检测出人血清中CMLC的含量,重复性好,灵敏度高,是临床诊断心肌梗塞的良好方法。

Objective To develop a sandwich ELISA method for detection of human cardiac myosin light chain（CMLC）,and primarily apply this method to diagnose acute myocardial infarction.Methods Two pairs of well-matched CMLC monoclonal antibodies were selected via the sandwich test and the measurement of the additive indices and affinity constants.The sandwich ELISA method was tried to measure the serum myosin CMLC of the patients.Results The best coating concentration of McAB was 5～7μg/mL.In the range of 1 to 50ng/mL,the CMLC antigen showed a good linearity in the standard curve.The serum CMLC concentration of the control group and patients were（0.875±0.162）（n=20） and（1.463±0.473）（n=30）,respectively.Conclusion The sandwich ELISA method can be used for detection of serum CMLC with good sensitivity and reproducibility,and is a good technique for the diagnosis of acute myocardiac infarction.