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| Establishment of the Micro-agglutination Test for Antibody Detection ofCitrobacter freundii from Sheep (Ovis aries) |
| LI Jing1,2, LIU Yi-Ting1,2, YANG Jin-Qian1,2, DING Lue-Xuan1,2, XUE Yun1,2, DING Ke1,2, SI Li-Fang1,2, ZHAO Zhan-Qin1,2* |
1 Key Lab of Animal Bacterial Infectious Disease Prevention, Control Technology, College of Animal Science, Technology, Henan University of Science, Techology, Luoyang 471003, China;
2 Key-Disciplines Lab of Safety of Environment, Animal Product, College of Animal Science, Technology, Henan University of Science, Techology, Luoyang 471003, China |
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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) byoptimizing the reaction conditions. The optimal antigen concentrations of inactivated whole-cell of C. freundiiLK-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 samplespositive for 8 common ovine pathogens. The antibodies in sheep could be detected at 14 d post vaccinationwith 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.
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Received: 26 July 2024
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Corresponding Authors:
*zhaozhanqin@126.com
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