Preparation and Application of Rabbit Polyclonal Antibodies against <i>Elizabethkingia meningosepticum</i> on Frogs

ZHANG Xinyan; ZENG Zhanzhuang; YE Xiaojun; ZHUO Yuchen; ZHONG Quanfu; YU Peijian

doi:10.19303/j.issn.1008-0384.2021.07.008

Volume 36 Issue 7

Jul. 2021

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Article Navigation > Fujian Journal of Agricultural Sciences > 2021 > 36(7): 785-792

ZHANG X Y, ZENG Z Z, YE X J, et al. Preparation and Application of Rabbit Polyclonal Antibodies against Elizabethkingia meningosepticum on Frogs [J]. Fujian Journal of Agricultural Sciences，2021，36（7）：785−792 doi: 10.19303/j.issn.1008-0384.2021.07.008

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ZHANG X Y, ZENG Z Z, YE X J, et al. Preparation and Application of Rabbit Polyclonal Antibodies against Elizabethkingia meningosepticum on Frogs [J]. Fujian Journal of Agricultural Sciences，2021，36（7）：785−792 doi: 10.19303/j.issn.1008-0384.2021.07.008

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Preparation and Application of Rabbit Polyclonal Antibodies against Elizabethkingia meningosepticum on Frogs

doi: 10.19303/j.issn.1008-0384.2021.07.008

The Freshwater Fisheries Research Institute of Fujian Province, Fuzhou, Fujian　350002, China

Received Date: 2021-04-15
Rev Recd Date: 2021-05-31

Available Online: 2021-07-13

Publish Date: 2021-07-28

Abstract

Abstract

Objective Rabbit polyclonal antibodies against Elizabethkingia meningosepticum (Em) were prepared to establish an indirect ELISA detection to trace the pathogenic attack in organs of frogs (Rana spp). Method The strain RsB1151018NA of Em from a diseased R. spinosa was isolated, formalin-inactivated, and injected into a rabbit for the serum preparation. An optimized indirect ELISA using the rabbit polyclonal antibodies was developed for tracing RsB1151018NA. The rabbit immunoglobulin G was purified on a protein A column. The purified antibodies were marked with FITC and further purified on a desalting column Sephadex G-25 for subsequently tracing the pathogen in organs of the inoculated frogs. Results The obtained rabbit polyclonal antibodies showed a titer up to 1 ∶ 5.12×10⁵, and 1 ∶ 1.6×10⁴ after purification. The indirect ELISA assay on the polyclonal antibody was specific and sensitive in detecting Em without cross-reaction on other bacteria. It had a detection limit of 1.0×10⁴ cfu·mL⁻¹. According to the in vivo tracing test, RsB1151018NA distributed throughout the entire frog body by blood circulation with the most serious infections in the kidney, eyes, and brain. Conclusion The optimized indirect ELISA detection of Em was rapid, sensitive, and specific in tracing the bacterial infection on R. spp. It was considered applicable for early diagnosis of the “crooked-head disease” and breeding of healthy frogs for food and medicine.
- Elizabethkingia meningosepticum,
- polyclonal antibodies,
- ELISA,
- detection,
- “crooked-head disease” of frogs

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