Gold(III) enhanced chemiluminescence immunoassay for detection of antibody against ApxIV of Actinobacillus pleuropneumoniae.
We report, for the first time, a chemiluminescence immunoassay (CLIA) method based on AuCl(4)(-)-enhanced luminol chemiluminescence (CL) reaction for the highly sensitive detection of ApxIV antibody of Actinobacillus pleuropneumoniae (APP). The AuCl(4)(-), which was the dissolution product of the gold nanoparticle-rabbit anti-pig IgG conjugate, served as an analyte in the CL reaction for the indirect measurement of antibody against ApxIV. The optimal condition of gold dissolution was composed of a 5.0 x 10(-2) M HCl, 1.5 x 10(-2) M NaCl, and 2.5 x 10(-4) M Br(2) solution. Under the optimal conditions, a good correlation between the relative CL photon counting and the dilution coefficient of serum was obtained in the dilution range of 1:160-1:40 000. Based on the analysis of clinical samples, the results indicated that CLIA had remarkable advantages in terms of reliability and practical use compared with indirect hemagglutination (IHA) and enzyme-linked immunosorbent assays (ELISA). The proposed method provided a new tool for the indirect determination of antibody against ApxIV in pig serum samples and showed great potential for numerous applications in immunoassays.
Hu
Dehong
D
College of Science, State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China. hyhan@mail.hzau.edu.cn.
Han
Heyou
H
Zhou
Rui
R
Dong
Fei
F
Bei
Weicheng
W
Jia
Fan
F
Chen
Huanchun
H
D016977:Actinobacillus pleuropneumoniae; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D001426:Bacterial Proteins / Q000276:immunology*; D006046:Gold; D007118:Immunoassay / Q000379:methods*; D008163:Luminescent Measurements; D036103:Nanotechnology; D011014:Pneumonia / Q000175:diagnosis*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis*
D000907:Antibodies, Bacterial; C122177:ApxIVA protein, Actinobacillus pleuropneumoniae; D001426:Bacterial Proteins; D006046:Gold