Development of Polyclonal Antibody-Based Indirect Enzyme-Linked Immunosorbent Assay for the Detection of Alicyclobacillus Strains in Apple Juice
Article first published online: 26 OCT 2012
© 2012 Institute of Food Technologists®
Journal of Food Science
Volume 77, Issue 11, pages M643–M649, November 2012
How to Cite
Wang, Z., Yue, T., Yuan, Y., Cai, R., Guo, C., Wang, X. and Niu, C. (2012), Development of Polyclonal Antibody-Based Indirect Enzyme-Linked Immunosorbent Assay for the Detection of Alicyclobacillus Strains in Apple Juice. Journal of Food Science, 77: M643–M649. doi: 10.1111/j.1750-3841.2012.02961.x
- Issue published online: 19 NOV 2012
- Article first published online: 26 OCT 2012
- MS 20120376 Submitted 3/10/2012, Accepted 8/25/2012.
- apple juice;
- indirect enzyme-linked immunosorbent assay;
- polyclonal antibody;
- Western blot
Abstract: A sort of specific polyclonal anti-Alicyclobacillus antibody was generated by immunizing New Zealand white rabbits, and a sensitive indirect enzyme-linked immunosorbent assay (ELISA) was developed for Alicyclobacillus detection in apple juice. A set of experimental parameters such as concentration of antigen, dilutions of the antibody and goat anti-rabbit IgG-horseradish peroxidase conjugate, selection of the blocking reagent, incubation time, and temperature was optimized. The cross-reactivity of the antibody was evaluated by ELISA and the result was consistent with Western blot analysis. The detection limit of the ELISA was about 105 colony forming units (CFU)/mL in apple juice samples. Samples were detected by ELISA and conventional culture method, and the ELISA results gave a good agreement with the results obtained by plating on Alicyclobacillus acidoterrestris medium agar. ELISA takes a total detection time of 6 to 7 h, which is less than the time of conventional techniques requiring more than 24 to 48 h. These results indicated that the established ELISA was a potential useful analytical method for detection of Alicyclobacillus in apple juice.