农业生物技术学报2025，Vol.33Issue(6)：1409-1416,8.DOI:10.3969/j.issn.1674-7968.2025.06.020
羊源弗氏柠檬酸杆菌微量凝集试验抗体检测方法的建立
Establishment of the Micro-agglutination Test for Antibody Detection of Citrobacter freundii from Sheep(Ovis aries)
摘要
Abstract
Ovine Citrobacter freundii was an emerging infectious disease affecting sheep(Ovis aries),in order to establish a rapid detection method for the disease,this study used the inactivated whole-cell of C.freundii LK-1 strain as the antigen,establishing an antibody detection method of micro-agglutination test(MAT)by optimizing the reaction conditions.The optimal antigen concentrations of inactivated whole-cell of C.freundii LK-1 strain were 2.0×109 cfu/mL and the incubation temperature was 37 ℃ for 7～9 h.The MAT was specificity for C.freundii of sheep,demonstrating negative reactivity(titer≤1∶2)against serum samples positive for 8 common ovine pathogens.The antibodies in sheep could be detected at 14 d post vaccination with inactivated C.freundii LK-1 vaccines.The values at day 21 and 35 were more than 1∶16 and 1∶64,respectively.The intra-assay coefficients of variation(CV)were consistently 0％,while the inter-assay CV ranged from 5.43％to 6.60％,indicating acceptable reproducibility.374 serum samples,which were detected by the developed MAT and the ELISA with the same antigen.The results of tests showed that the total positive rates were 9.9％and 10.7％,respectively.The coincidence of the positive and negative results was good(Kappa value was 0.897),showing that the sheep were infected by C.freundii in a certain area of Henan Province.This study provides a basis for vaccine antibody evaluation and epidemiological investigation of ovine C.frendii.关键词
羊/弗氏柠檬酸杆菌/微量凝集试验/抗体Key words
Sheep/Citrobacter freundii/Microagglutination test/Antibody分类
农业科技引用本文复制引用
李静,刘艺婷,杨金钱,丁略烜,薛云,丁轲,司丽芳,赵战勤..羊源弗氏柠檬酸杆菌微量凝集试验抗体检测方法的建立[J].农业生物技术学报,2025,33(6):1409-1416,8.基金项目
国家自然科学基金(32072899 （）
U1704117) （）
