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1. 山东大学控制科学与工程学院 生物医学工程系,山东 济南,250061
2. 齐鲁工业大学 机械与汽车工程学院,山东 济南,250353
3. 山东大学 附属济南市中心医院,山东 济南,250013
4. 山东大学 附属山东省立医院,山东 济南,250021
Received:12 December 2013,
Revised:28 January 2014,
Published:03 April 2014
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刘剑, 张丽丽, 汪运山等. 增强型酶催化发光免疫分析法高灵敏测定血清中心肌肌钙蛋白Ⅰ[J]. 发光学报, 2014,35(4): 506-510
LIU Jian, ZHANG Li-li, WANG Yun-shan etc. A Highly Sensitive Determination of Cardiac Troponin Ⅰ in Serum Samples by Enhanced Enzymatic Chemiluminescence Immunoassay[J]. Chinese Journal of Luminescence, 2014,35(4): 506-510
刘剑, 张丽丽, 汪运山等. 增强型酶催化发光免疫分析法高灵敏测定血清中心肌肌钙蛋白Ⅰ[J]. 发光学报, 2014,35(4): 506-510 DOI: 10.3788/fgxb20143504.0506.
LIU Jian, ZHANG Li-li, WANG Yun-shan etc. A Highly Sensitive Determination of Cardiac Troponin Ⅰ in Serum Samples by Enhanced Enzymatic Chemiluminescence Immunoassay[J]. Chinese Journal of Luminescence, 2014,35(4): 506-510 DOI: 10.3788/fgxb20143504.0506.
基于辣根过氧化物酶(HRP)催化 H
2
O
2
-Luminol系统,对一种新型酚类衍生物 4-(1,2,4-三氮唑-1-基)苯酚对化学发光的增强作用进行了研究。用HRP标记急性心肌梗死标志物心肌肌钙蛋白Ⅰ(cTnⅠ)单克隆抗体,通过cTnⅠ的双抗体夹心免疫反应,建立了简单、灵敏和快速检测人血清中的cTnⅠ含量的增强型酶发光免疫分析方法。实验结果表明,cTnⅠ在1.2 ~ 24 ng/mL浓度范围内与增强型发光强度具有良好的线性关系(
R
=0.99),变异系数(
n
= 8)为 4.7%。最后,使用该方法对人血清样品中的cTnⅠ含量进行测定,测试结果具有较好的稳定性和精度,能够满足临床检测要求。
Based on the catalyzed oxidation of horseradish peroxidase (HRP) to the hydrogen peroxide and luminol system
a novel p-phenol derivative
4-(1
2
4-triazol-1-yl)phenol (TP) was studied to enhance the chemiluminescence. With HRP labeled cardiac troponin Ⅰ (cTn Ⅰ) monoclonal antibody
a simple
sensitive
and rapid enzyme chemiluminescence immunoassay method enhanced by TP for the determination of cTn Ⅰ in human serum samples had been developed by "sandwich type" immune response.The experimental results show that there is a very good linear correlation between the enhanced enzymatic chemiluminescence intensity and the amount of cTn Ⅰ in the range of 1.2 ~ 24 ng/mL (
R
= 0.99)
and the coefficient of variation (
n
=8) is 4.7%. The proposed enhanced enzymatic chemiluminescence immunoassay method had been used successfully for the determination of cTn Ⅰ in human serum.
Sarapultsev P, Chupakhin O, Sarapultsev A, et al. New insights in to the treatment of myocardial infarction[J]. Int. J. Exp. Pathol., 2012, 93(1):18-23. [2] The Joint European Society of Cardiology/American College Cardiology Committee. Myocardial infarction redefinedA consensus document of the Joint European Society of Cardiology/American College Cardiology Committee for the redefined of myocardial infarction[J]. J. Am. Coll. Cardial, 2000, 36(3):959-969. [3] Ilva T, Lund J, Porela P, et al. Early markers of myocardial injury: cTnⅠ is enough[J]. Clin. Chim. Acta, 2009, 400(1-2):82-85. [4] Song S Y, Han Y D, Kim K, et al. A fluoro-microbead guiding chip for simple and quantifiable immunoassay of cardiac troponin Ⅰ(cTnⅠ)[J]. Biosens. Bioelectron., 2011, 26(9):3818-3824. [5] Geri G, Mongardon N, Dumas F, et al. Diagnosis performance of high sensitivity troponin assay in out-of-hospital cardiac arrest patients[J]. Int. J. Cardiol., 2013, 169(6):449-454. [6] Lee J, Lee Y, Park J Y, et al. A centrifugally actuated point-of-care testing system for the surface acoustic wave immunosensing of cardiac troponin I[J]. Sens. Actuat. B: Chem., 2013, 178:19-25. [7] Cummins B, Russell G J, Chandler S T, et al. Uptake of radioiodinated cardiac specific troponin-I antibodies in myocardial infarction[J]. Cardiovasc Res., 1990, 24(4):317-327. [8] Iqbal A, Ateeq N. Effect of processing on the detectability of peanut protein by ELISA[J]. Food Chem., 2013, 141(3):1651-1654. [9] Marine W, Florence C. Evaluation of ELISA for detection of rabies antibodies in domestic carnivores[J]. J. Virol. Methods, 2012, 179(1):166-175. [10] Liu J, Zhang L L, Wang Y S, et al. An improved portable biosensing system based on enzymatic chemiluminescence and magnetic immunoassay for biological compound detection[J]. Measurement, 2014, 47:200-206. [11] Zhao J W, Chen Y N, Lu S Z, Key technologies of chemiluminescence immunoassay analyzer[J]. Chin. J. Lumin.(发光学报), 2012, 33(12):1381-1388 (in Chinese). [12] Xiao Q, Li H, Lin J M. Development of a highly sensitive magnetic particle-based chemiluminescence enzyme immunoassay for thyroid stimulating hormone and comparison with two other immunoassays[J]. Clin. Chim. Acta, 2010, 411(15-16):1151-1153. [13] Liu Z J, Song C J, Li Y M, et al. Development of highly sensitive chemiluminescence enzyme immunoassay based on the anti-recombinant H-C subunit of botulinum neurotoxin type a monoclonal antibodies[J]. Anal. Chim. Acta, 2012, 735:23-30. [14] Yang X Y, Guo Y S, Mei Z H, Chemiluminescent determination of H2O2 using 4-(1, 2, 4-triazol-1-yl)phenol as an enhancer based on the immobilization of horseradish peroxidase onto magnetic beads[J]. Anal. Biochem., 2009, 393(1):56-61. [15] Tian D, Duan C, Wang W, et al. Sandwich-type electrochemiluminescence immunosensor based on N-(aminobutyl)-N-ethylisoluminol labeling and gold nanoparticle amplification[J]. Talanta, 2009, 78(2):399-404.
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