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Detection of anticardiolipin antibody IgG by time-resolved fluoroimmunoassay

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Abstract

In an effort to improve the quantitative detection of anticardiolipin antibodies (aCL) IgG so as to classify patients correctly as antiphospholipid syndrome (APS) positive, we developed a new immunoassay based on a sandwich time-resolved fluoroimmunoassay (TRFIA) using the complex of cardiolipin plus bovine β2GPI as antigen and Eu3+-labeled rabbit antihuman IgG as conjugate. The precision, sensitivity, specificity, and stability of the assay were evaluated, and comparison with the classical ELISA was also made. The aCL IgG TRFIA kit we established had a wider detectable range than three commercial ELISA ones from different manufacturers when a specimen was diluted, with strong positive result from 1:12.5 to 1:204,800. The average intra-assay and inter-assay CVs detected by the aCL IgG TRFIA was 3.14 and 3.70 %, respectively. The sensitivity was 0.1 GPL U/ml, and the clinical diagnostic specificity was 98 %. The established assay kit also behaved better in stability than the commercial ELISA ones. Additionally, the immunoassay we established correlated well with the ELISA, and the correlation coefficient was 0.975. We thus conclude that the TRFIA we developed for aCL IgG detection gives promise to a more sensitive and reliable diagnosis of APS and has potential value for large-scale screening programs.

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Acknowledgments

This work was supported by the Directive Planning Project of Wuxi Science and Technology Bureau (CSE01010), China.

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Authors and Affiliations

  1. Affiliated Wuxi People’s Hospital of Nanjing Medical University, Wuxi, 214023, China

    Yan Ye, Zhigang Hu, Jie Liu, Guoqian Chen, Yaohong Zhou & Lei Yu

  2. Department of Clinical Laboratory, Affiliated Wuxi People’s Hospital of Nanjing Medical University, Wuxi, 214023, China

    Zhigang Hu

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  1. Yan Ye
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Correspondence to Zhigang Hu.

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Ye, Y., Hu, Z., Liu, J. et al. Detection of anticardiolipin antibody IgG by time-resolved fluoroimmunoassay. Clin Rheumatol 31, 1339–1345 (2012). https://doi.org/10.1007/s10067-012-2020-6

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  • DOI: https://doi.org/10.1007/s10067-012-2020-6

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