人心肌特异表达基因Nelin蛋白的血清水平测定及其意义

目的探讨Nelin蛋白在心血管病患者和正常对照血清表达水平的差别及其意义。方法使用自制的一株Nelin单克隆抗体(MA-H409H5),建立ELISA法标准曲线。通过检测灵敏度(最小检出量)、精确度(批内、批间变异系数)、回收率等以优化实验反应条件,并以此方法测定了580例人血清Nelin蛋白浓度。结果直接ELISA法检测Nelin蛋白的最小检出量为7.8ng/ml,标准曲线在7.8～250.0ng/ml范围内线性良好(r0.99)。精确度测定:平均批内变异系数为4.78%,平均批间变异系数为9.56%。测得回收率为(100±5)%。580例人血清中均可检测到Nelin蛋白的表达,其中正常对照组为(9.34±3.07)μg/ml,冠心病组为(8.57±2.76)μg/ml,该两组Nelin蛋白含量相比有显著性差异(P0.05),而其他各心脏病组与正常组相比均无显著性差异。结论成功建立了人血清Nelin蛋白的ELISA测定法,该方法敏感、特异、可靠。正常对照组和各心血管病患者组均有Nelin蛋白表达,但冠心病组明显低于正常对照组,提示Nelin基因的表达水平可能与冠心病有一定的关系。

Detection of the Serum Levels of Human Cardiac-specific Gene Nelin and Its Clinical Significance

Objective Exploring the serum levels of Nelin protein, a novel cardiac-specificprotein, and comparing the difference in the normal control and patients with heart diseases. Methods The standard curve was plotted using the specific monoclonal antibody （MA-H409 H5 ） which was raised in our laboratory. We determined the assay sensitivity（ the detection limits） , precision （coefficient of variation for inner-batch and inter-batch ） and recovery. The serum levels of Nelin protein with 580 samples were detected by the optimized ELISA method. Results The standard curve is linearly related from 7.8 to 250.0 ng/ml （r〉0.99）. The detection limit of Nelin protein was 7.8 ng/ml. Coefficients of variation for inner-batch and inter-batch were 4.78 % and 9.56 % respectively. The recovery was（ 100±5）%. The concentration of Nelin in serum was measured in total 580 samples. The serum level of Nelin protein was（9.34±3.07）μg/ml in normal control, and （8.57±2.76） μg/ ml in patients of coronary heart disease. There is significant difference between the two groups（P〈 0. 05）, but no significant correlation can be found within other patients and control. Conclusion Detection method for human serum Nelin is successfully established. The assay results were specific, sensitive and reliable. We have proved that Nelin protein in serum exists in normal and several patients groups. Especially, the protein level of coronary heart disease group is significantly lower than that in the control group. The study indicates that the expression level of Nelin, to some degree, is predictor of coronary heart diseases.