title: Development of polyclonal antibody-based indirect enzyme-linked immunosorbent assay for the detection of Alicyclobacillus strains in apple juice.
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 10(5)  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.
authors.affiliation: College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi, 712100, China.
authors.forename: Zhouli
authors.initials: Z
authors.lastname: Wang
authors.forename: Tianli
authors.initials: T
authors.lastname: Yue
authors.forename: Yahong
authors.initials: Y
authors.lastname: Yuan
authors.forename: Rui
authors.initials: R
authors.lastname: Cai
authors.forename: Caixia
authors.initials: C
authors.lastname: Guo
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Chen
authors.initials: C
authors.lastname: Niu
mesh_terms: D056450:Alicyclobacillus / Q000302:isolation & purification*; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D001628:Beverages; D015153:Blotting, Western; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D006735:Horseradish Peroxidase / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry; D008297:Male; D027845:Malus / Q000382:microbiology*; D011817:Rabbits; D012680:Sensitivity and Specificity
chemical_list: D000906:Antibodies; C051538:IGG-horseradish peroxidase; D007074:Immunoglobulin G; D006735:Horseradish Peroxidase