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间接ELISA测定人血IgM型抗-A抗体方法的建立及应用
Establishment and application of an indirect ELISA for determining IgM anti-A antibodies in human plasma
- 第三军医大学学报 2018年40卷第7期 页码：556-560
- 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：
[赵树铭，E-mail:shumingzhao123@qq.com ZHAO Shuming, E-mail:shumingzhao123@qq.com
- 基金信息：全军“十二五”计划专项课题(ASW11J007-05);第三军医大学西南医院院管课题(SWH2014SX02);国家自然科学基金面上项目(81570167)
DOI：10.16016/j.1000-5404.201711078
中图分类号： R392-33;R392.11;R457.11收稿：2017-11-10，
修回：2018-01-02，
纸质出版：2018-04-15
- 稿件说明：
移动端阅览
宋敏, 范娅涵, 王世春, 等. 间接ELISA测定人血IgM型抗-A抗体方法的建立及应用[J]. 第三军医大学学报, 2018,40(7):556-560.
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.
宋敏, 范娅涵, 王世春, 等. 间接ELISA测定人血IgM型抗-A抗体方法的建立及应用[J]. 第三军医大学学报, 2018,40(7):556-560. DOI： 10.16016/j.1000-5404.201711078.
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
2
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
2
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
2
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
D
(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(
r
=0.71，
P
<
0.01)。
结论
2
成功建立了人血IgM型抗-A抗体定量检测方法。
Abstract
Objective
2
To develop an indirect enzyme linked immunosorbent assay (ELISA) for rapid and quantitative detection of IgM anti-A antibodies in human plasma.
Methods
2
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
2
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
D
(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 (
r
=0.71
P
<
0.01).
Conclusion
2
An indirect ELISA is developed successfully for quantitative detection of IgM anti-A antibodies.
关键词
Keywords
references
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