Establishment and application of an indirect ELISA for determining IgM anti-A antibodies in human plasma

Min SONG; Yahan FAN; Shichun WANG; Qiang ZHANG; Ting XU; Tianlun JIANG; Shuming ZHAO

doi:10.16016/j.1000-5404.201711078

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Establishment and application of an indirect ELISA for determining IgM anti-A antibodies in human plasma

Basic Medicine | 更新时间：2026-01-09

- Establishment and application of an indirect ELISA for determining IgM anti-A antibodies in human plasma
- Journal of Third Military Medical University Vol. 40, Issue 7, Pages: 556-560(2018)
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)第一附属医院输血科
- 作者简介：
- 基金信息：
  
  the Special Project of the "Twelfth Five-year Plan" for Medical Science Development of PLA(ASW11J007-05);the Project of Southwest Hospital of Third Military Medical University(SWH2014SX02);the General Program of National Natural Science Foundation of China(81570167)
- DOI：10.16016/j.1000-5404.201711078
  CLC： R392-33;R392.11;R457.11
- Received：10 November 2017，
  
  Revised：2018-01-02，
  
  Published：15 April 2018
- 稿件说明：
移动端阅览
Min SONG, Yahan FAN, Shichun WANG, et al. Establishment and application of an indirect ELISA for determining IgM anti-A antibodies in human plasma[J]. Journal of Third Military Medical University, 2018, 40(7): 556-560.
DOI：

Min SONG, Yahan FAN, Shichun WANG, et al. Establishment and application of an indirect ELISA for determining IgM anti-A antibodies in human plasma[J]. Journal of Third Military Medical University, 2018, 40(7): 556-560. DOI： 10.16016/j.1000-5404.201711078.

摘要

目的

建立间接酶联免疫吸附实验(enzyme linked immunosorbent assay，ELISA)检测方法，快速定量检测人血IgM抗-A抗体。

方法

通过制备A型红细胞膜蛋白作为捕获抗原，采用棋盘滴定实验筛选出最佳抗原抗体工作浓度，建立人血IgM型抗-A抗体定量检测方法。

结果

所构建的间接ELISA定量检测方法检测下限为0.02 μg/mL；批内变异系数及批间变异系数均小于10%；通过ROC曲线分析，当临界

(450)为0.259时，ROC曲线下面积最大，为0.947，检测灵敏度为87.7%，特

异性为100%。采用本研究自建的ELISA检测57例B型血浆标本的IgM型抗-A抗体浓度为(1.71±1.95)μg/mL，定量结果与经典半定量效价评分呈正相关(

=0.71，

0.01)。

结论

成功建立了人血IgM型抗-A抗体定量检测方法。

Abstract

Objective

To develop an indirect enzyme linked immunosorbent assay (ELISA) for rapid and quantitative detection of IgM anti-A antibodies in human plasma.

Methods

An in-house indirect ELISA was developed using the ghost protein of group A erythrocyte as coating antigen. The optimal combination of antigens and antibodies was selected by checkerboard titration. The methodological performance was evaluated

and the plasma concentrations of IgM anti-A antibodies of clinical samples were determined.

Results

The detection limit of the in-house indirect ELISA was 0.02 μg/mL. The coefficient of variation for inner-batch and inter-batch were both less than 10%. According to the receiver operating characteristic (ROC) curve analysis

the largest area under the curve accounted for 0.947 when the cut-off value at

(450) was 0.259

and the detection sensitivity and specificity were 87.7% and 100% respectively. The plasma level of IgM anti-A antibodies in 57 samples of type B plasma was 1.71±1.95 μg/mL. The quantitative results were positively correlated to the titer scores from the classical semi-quantitative detection of IgM anti-A antibodies (

=0.71

0.01).

Conclusion

An indirect ELISA is developed successfully for quantitative detection of IgM anti-A antibodies.

关键词

Keywords

references

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