[Development of time-resolved fluoroimmunoassay kit for detection of IgM antibodies against hepatitis B core antigen].
OBJECTIVE
To prepare a time-resolved fluoroimmunoassay (TRFIA) kit for clinical detection of IgM antibodies to hepatitis B core antigen (HBc).


METHODS
Immunocapture method was used to develop the TRFIA kit for detection of the anti-HBc IgM antibodies, and the precision, cross-reactivity and sensitivity of the kit were tested with the clinical serum samples.


RESULTS
The intra- and inter-assay coefficients of variation of the TRFIA kit were 4.8%-7.2% and 7.5%-8.6%, respectively, and no cross-reactivity with anti-HBs, anti-HBc-IgG or anti-HBe was found. Comparison of the results of the TRFIA kit and enzyme-linked immunosorbent assay (ELISA) demonstrated greater sensitivity of the kit than ELISA in detecting the anti-HBc IgM antibodies in 584 serum samples. According to the detection results in 300 serum samples from healthy donors, the cutoff value of the TRFIA kit was 4.5 times of the fluorescence value of the negative control.


CONCLUSIONS
This TRFIA kit for detecting anti-HBc IgM antibodies meets the demand for clinical application and can replace the ELISA kits.
Hu
Ni-ya
NY
College of Biotechnology, Southern Medical University, Guangzhou 510515, China. huniya@sina.com
Dong
Zhi-ning
ZN
Zhou
Zhuo-long
ZL
Li
Ming
M
Wu
Ying-song
YS
D015200:Fluoroimmunoassay / Q000379:methods*; D006509:Hepatitis B / Q000276:immunology / Q000821:virology*; D006510:Hepatitis B Antibodies / Q000097:blood*; D006512:Hepatitis B Core Antigens / Q000276:immunology*; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity
D006510:Hepatitis B Antibodies; D006512:Hepatitis B Core Antigens; D007075:Immunoglobulin M; D011933:Reagent Kits, Diagnostic