Abstract:

Objective: To explore the effect of storage temperature on the stability of the laboratory-fabricated enzyme linked
immunosorbent assay (ELISA) kit for AFB1 detection. Methods: Changes in OD value of blank control wells, IC20, IC50 and
IC80 (the concentrations required for 20%, 50% and 80% inhibition, respectively) and RSD were examined upon addition of
different concentrations of AFB1 when stored at 4 or 37 ℃ and the R² values of the standard curves at different storage times
were investigated. Results: On the 270th day of storage at 4 ℃, the OD value of blank control wells varied from 1.129 to 1.777
and the IC20, IC50 and IC80 were 0.05, 0.27 ng/mL and 0.92 ng/mL, with RSD of 27.36%, 27.58% and 19.82%, respectively. The
standard curves developed at different time points during the 270 d storage period exhibited good linearity (R² greater than 0.97).
The OD value of blank control wells varied from 1.200 to 1.809 over a storage period of 13 d under the accelerated conditions
at 37 ℃ and the IC20, IC50 and IC80 were 0.06, 0.25 ng/mL and 1.181 ng/mL with RSD of 27.71%, 25.66% and 21.89%,
respectively. Similarly, the standard curves developed at different time points during the 270 d storage period also showed good
linearity (R² greater than 0.98). Conclusion: The OD value of blank control wells in the ELISA kit remains above 1.0 regardless
of storage at 4 ℃ for 270 d or 37 ℃ for 13 d and all indexes examined can meet the requirements for in situ detection.

Key words: aflatoxin B1 (AFB1), enzyme linked immunosorbent assay (ELISA) kit, stability