Detection of anticardiolipin antibody IgG by time-resolved fluoroimmunoassay.
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 β(2)GPI as antigen and Eu(3+)-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.
Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi 214023, China.
Yan
Y
Ye
Zhigang
Z
Hu
Jie
J
Liu
Guoqian
G
Chen
Yaohong
Y
Zhou
Lei
L
Yu
D000818:Animals; D017153:Antibodies, Anticardiolipin / Q000032:analysis* / Q000276:immunology; D016736:Antiphospholipid Syndrome / Q000175:diagnosis* / Q000276:immunology; D002138:Calibration; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015200:Fluoroimmunoassay / Q000379:methods*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000737:chemistry*; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D017153:Antibodies, Anticardiolipin; D007074:Immunoglobulin G