A Highly Sensitive Chemiluminescence Immunoassay Technique for Assaying TSH in Human Serum

QI Su-wen; ZHANG Hui-sheng; LIU Xin-yu; LI Qiao-liang

doi:10.3788/fgxb20133408.1089

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A Highly Sensitive Chemiluminescence Immunoassay Technique for Assaying TSH in Human Serum

更新时间：2020-08-12

- A Highly Sensitive Chemiluminescence Immunoassay Technique for Assaying TSH in Human Serum
- Chinese Journal of Luminescence Vol. 34, Issue 8, Pages: 1089-1095(2013)
- 作者机构：
  
  深圳大学医学院生物医学工程系,广东深圳,518060
- 作者简介：
- 基金信息：
- DOI：10.3788/fgxb20133408.1089
  CLC： R318.15
- Received：16 May 2013，
  
  Revised：17 June 2013，
  
  Published：10 August 2013
- 稿件说明：
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齐素文, 张会生, 刘昕宇, 李乔亮. 高灵敏度化学发光免疫法检测人血清中促甲状腺激素[J]. 发光学报, 2013,34(8): 1089-1095

QI Su-wen, ZHANG Hui-sheng, LIU Xin-yu, LI Qiao-liang. A Highly Sensitive Chemiluminescence Immunoassay Technique for Assaying TSH in Human Serum[J]. Chinese Journal of Luminescence, 2013,34(8): 1089-1095
齐素文, 张会生, 刘昕宇, 李乔亮. 高灵敏度化学发光免疫法检测人血清中促甲状腺激素[J]. 发光学报, 2013,34(8): 1089-1095 DOI： 10.3788/fgxb20133408.1089.

QI Su-wen, ZHANG Hui-sheng, LIU Xin-yu, LI Qiao-liang. A Highly Sensitive Chemiluminescence Immunoassay Technique for Assaying TSH in Human Serum[J]. Chinese Journal of Luminescence, 2013,34(8): 1089-1095 DOI： 10.3788/fgxb20133408.1089.

摘要

建立了一种高灵敏度检测促甲状腺激素(TSH)的化学发光体系。将异鲁米诺(ABEI)和异硫氰酸荧光素(FITC)分别标记TSH的单克隆抗体

与待测TSH通过夹心法进行免疫反应形成抗原抗体复合物之后

加入抗-FITC抗体包被的磁珠

在磁场作用下加入缓冲液进行清洗

分离非特异性物质

加入发光底物NaOH和H

检测发光强度(RLU)。本方法的灵敏度为0.01 IU/mL

批内和批间变异系数均在10%之内

回收率在96%~103%之间。使用本方法与其他检测系统进行比对

相关系数为0.993 7。实验结果表明

基于ABEI发光检测TSH的化学方法灵敏度高、结果可靠

具有较高的应用价值。

Abstract

A highly sensitive magnetic particle-based chemiluminescence immunoassay (CLIA) technique for assaying thyroid stimulating hormone (TSH) in serum was described. Fluorescein isothiocyanate (FITC) and N-(aminobutyl)-N-(ethylisoluminol) (ABEI) were used to label two different monoclonal antibodies of anti-TSH. Both of the labeled antibodies combined with TSH to form a sandwiched immunoreaction that was monitored by chemiluminescence (CL) detection. The magnetic particles (MPs) that were coated with anti-FITC antibody served as both the solid phase and the separator. The method showed high sensitivty with a detection limit of 0.01 IU/mL. The coefficient of variation of intra-assay imprecision and the inter-assay imprecision were less than 10%. The average recoveries were 96%~103%. A correlation analysis against an established automated assay generated a correlation coefficient of 0.993 7. The proposed method demonstrates an acceptable performance for quantifying serum TSH and is suitable for the clinical lab.

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references

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