人血清肌红蛋白含量的胶体金免疫比色法初步研究
|
摘要
目的初步建立一种基于胶体金均相免疫比色技术定量检测血清中肌红蛋白(myoglobin,Myo)的方法。方法制备直径80nm左右胶体金颗粒并标记Myo抗体,利用胶体金标记抗体与样本中Myo抗原结合后导致胶体金溶液颜色变化且变化程度与结合量呈比的原理,通过检测结合前后540nm与660nm处吸光度的变化进行肌红蛋白的定量。结果制备的胶体金颗粒液相分布均匀,胶体金颗粒标记抗体呈现均相分布,符合检测实验要求。建立了胶体金均相免疫比色检测肌红蛋白的方法,与化学发光方法的检测结果符合率达95. 10%。结论初步完成了肌红蛋白的胶体金免疫比色法的建立,与成熟的化学发光方法检测结果具有一定的符合性,有必要进行更深一步的研究。
Objective To establish a colloidal gold colorimetric assay method for the detection of myoglobin(Myo)in serum. Methods The colloid gold particles with diameter of 80 nm were prepared and labeled with Myo antibody. According to the mechanism that the color of the colloid gold solution would change upon the binding with antigen and the degree of change is proportionated to the amount of bindings of the colloid gold labeled antibody to the Myo antigen in the sample,the quantitative of the myoglobin was detected by measuring the changes of the absorbance at 540 nm and 660 nm before and after the combination. Results The colloidal gold particles were well distributed,and the prepared antibodies labeled colloidal gold particles were homogeneously distributed,which were suitable for the requirements of the test. In the preliminary analysis of the clinical specimens,compared with the chemiluminescence method,the coincidence rate was 95. 10%.Conclusion A colloidal gold colorimetric assay method for myoglobin in the human serum was established preliminarily,with the results consistent with those obtained by chemiluminescence method,and it can be further applied to the clinical detection.
Objective To establish a colloidal gold colorimetric assay method for the detection of myoglobin(Myo)in serum. Methods The colloid gold particles with diameter of 80 nm were prepared and labeled with Myo antibody. According to the mechanism that the color of the colloid gold solution would change upon the binding with antigen and the degree of change is proportionated to the amount of bindings of the colloid gold labeled antibody to the Myo antigen in the sample,the quantitative of the myoglobin was detected by measuring the changes of the absorbance at 540 nm and 660 nm before and after the combination. Results The colloidal gold particles were well distributed,and the prepared antibodies labeled colloidal gold particles were homogeneously distributed,which were suitable for the requirements of the test. In the preliminary analysis of the clinical specimens,compared with the chemiluminescence method,the coincidence rate was 95. 10%.Conclusion A colloidal gold colorimetric assay method for myoglobin in the human serum was established preliminarily,with the results consistent with those obtained by chemiluminescence method,and it can be further applied to the clinical detection.
引文
[1]陈玲玲.急性心肌梗死患者血清肌红蛋白肌钙蛋白以及超敏C-反应蛋白含量动态监测的临床诊断意义[J].安徽医学,2012,33(9):1224-1226.
[2]黄磊,卓玲,赵越,等.肌红蛋白的代谢及其在心血管系统中的作用[J].心血管病学进展,2017,38(4),451-454.
[3]杨靖,宋谦.血清肌红蛋白、肌钙蛋白I及炎症性因子水平检测对心肌梗死的临床诊断意义[J].检验医学与临床,2016,13(16):2322-2323.
[4]尚红,王毓三,申子瑜.全国临床检验操作规程[M].第4版.北京:人民卫生出版社,2015.
[5]LEUVERING J H,THAL P J,van der WAART M,et al. Sol particle immunoassay(spia)[J]. J Immunoassay,1980,1:77-91.
[6]程黎明,朱晓华,朱耀武.液相免疫胶体金技术用于检测血浆半胱氨酸蛋白酶抑制剂C的方法学评价[J].医学检验,2010,25(3):214-218.
[7]王会中.一种肌红蛋白检测试剂及其制备方法[P].中国专利:201410648653. 0,2015-03-04.
[8]王会中,徐荣.定量免疫分析技术的应用现状与展望[J].中华检验医学杂志,2017,40(6):478-480.
[9]张莹,冯海,吴秋平,等.胶体金免疫层析技术的研究进展[J].现代生物医学进展,2015(12):2349-2351.
[10]戴婉如,陈发林,吴来州.免疫胶体金技术测定血清胱抑素C及其对早期肾损害的诊断价值[J].检验医学与临床,2009,6(5):336-337.
[11]KYKSE ANDERSON J,SCHMIDT C,NORDIN G,et al. Determined by a rapid,automated particle enhanced turbidimetric method,is a better marder than serum creatinine for glomerular filtration rate[J]. Clin Chem,1994,40:1921-1926.
[12]FINNEY H,NEWMAN D J,GRUBER W,et al. Initial evaluation of cystatin C measurement by particle enhanced immunonephelometry on the Behring nephelometry systems(BNA,BN11)[J]. Clin Chem,1997,43:1016-1022.
[13]肖勤,林金明.化学发光免疫分析新进展[J].上海医学检验杂志,2003,18(1):6-8.
[2]黄磊,卓玲,赵越,等.肌红蛋白的代谢及其在心血管系统中的作用[J].心血管病学进展,2017,38(4),451-454.
[3]杨靖,宋谦.血清肌红蛋白、肌钙蛋白I及炎症性因子水平检测对心肌梗死的临床诊断意义[J].检验医学与临床,2016,13(16):2322-2323.
[4]尚红,王毓三,申子瑜.全国临床检验操作规程[M].第4版.北京:人民卫生出版社,2015.
[5]LEUVERING J H,THAL P J,van der WAART M,et al. Sol particle immunoassay(spia)[J]. J Immunoassay,1980,1:77-91.
[6]程黎明,朱晓华,朱耀武.液相免疫胶体金技术用于检测血浆半胱氨酸蛋白酶抑制剂C的方法学评价[J].医学检验,2010,25(3):214-218.
[7]王会中.一种肌红蛋白检测试剂及其制备方法[P].中国专利:201410648653. 0,2015-03-04.
[8]王会中,徐荣.定量免疫分析技术的应用现状与展望[J].中华检验医学杂志,2017,40(6):478-480.
[9]张莹,冯海,吴秋平,等.胶体金免疫层析技术的研究进展[J].现代生物医学进展,2015(12):2349-2351.
[10]戴婉如,陈发林,吴来州.免疫胶体金技术测定血清胱抑素C及其对早期肾损害的诊断价值[J].检验医学与临床,2009,6(5):336-337.
[11]KYKSE ANDERSON J,SCHMIDT C,NORDIN G,et al. Determined by a rapid,automated particle enhanced turbidimetric method,is a better marder than serum creatinine for glomerular filtration rate[J]. Clin Chem,1994,40:1921-1926.
[12]FINNEY H,NEWMAN D J,GRUBER W,et al. Initial evaluation of cystatin C measurement by particle enhanced immunonephelometry on the Behring nephelometry systems(BNA,BN11)[J]. Clin Chem,1997,43:1016-1022.
[13]肖勤,林金明.化学发光免疫分析新进展[J].上海医学检验杂志,2003,18(1):6-8.