[Preparation of time-resolved fluoroimmunoassay kit for human alpha-fetoprotein detection].
OBJECTIVE
To prepare time-resolved fluoroimmunoassay (TRFIA) kit for detecting human alpha-fetoprotein (hAFP).


METHODS
Sandwich TRFIA kit for hAFP detection was developed using monoclonal antibody of anti-AFP.


RESULTS
AFP-TRFIA kit was capable of detecting AFP within the range of 1-1 000 U/ml with sensitivity of 0.17 U/ml and without cross-reactivity with CEA, CA12-5, CA15-3, or CA19-9. The intra- and inter-assay coefficients of variation were (3.3-5.9)% and (3.7-6.5)%, respectively. The prepared AFP-TRFIA reagent could withstand preservation at 4 degrees celsius; for 1 year and at 37 degrees celsius; for 7 days with the cutoff value of 12 U/ml in healthy subjects (n=426). The correlation coefficient of the detection results between this kit and commercially available AFP kit (Wallac OY, Finland) for 60 blood samples was 0.995.


CONCLUSIONS
The prepared TRFIA kit for hAFP detection meets the demand of clinical application with good sensitivity, precision, specificity, stability and accuracy.
Hang
Jian-Feng
JF
Institute of Tropical Medicine, Southern Medical University, Guangzhou 510515, China. hangjianfeng_ren@163.com
Wu
Ying-Song
YS
Yu
Wei-Hong
WH
Xu
Wei-Wen
WW
Wang
Cong-Rong
CR
Qiu
Yu-Rong
YR
Li
Ming
M
D015200:Fluoroimmunoassay / Q000295:instrumentation*; D006801:Humans; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013997:Time Factors; D000509:alpha-Fetoproteins / Q000032:analysis*
D011933:Reagent Kits, Diagnostic; D000509:alpha-Fetoproteins