Preparation of monoclonal antibodies against Bc48 and development of a rapid detection assay for infection with Babesia caballi in China.
Abstract: Babesia caballi (Nuttal, 1910) is one of the causative agents of equine piroplasmosis which causes economic losses to horse industry in China. There is an urgent need for rapid detection method for B. caballi infection in Xinjiang Province, China. To prepare monoclonal antibodies (mAbs) against Bc48 gene of B. caballi (Xinjiang local strains) and establish colloidal gold-immunochromatographic (ICT) assay for diagnosis of the disease, recombinant Bc48 was expressed and purified from Escherichia coli. With purified Bc48 as immunogen in mice, three hybridoma cells named 11F4, 1H2 and 7F4 secreting mAbs against Bc48 of B. caballi were obtained, which showed strong reaction with recombinant Bc48 and Bc48 gene transfected cells. Furthermore, colloidal gold labelled ICT assay based on purified Bc48 recombinant antigen and its mAb was developed. The ICT assay showed high sensitivity and specificity and no cross-reaction with Theileria equi (Laveran, 1901). Total of 56 horse serum samples collected from Xinjiang were tested by ICT and compared with the detection by commercial ELISA kit. The results showed that 32 out of 56 serum samples were positive by ICT and 33 were positive by ELISA. ICT assay had high coincidence (98%) to the reference ELISA kit. mAbs and ICT developed in this study could be provided as an efficient diagnosis tool for infection with B. caballi in horse in Xinjiang area.
Publication Date: 2019-05-13 PubMed ID: 31124789DOI: 10.14411/fp.2019.005Google Scholar: Lookup
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Summary
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This study summarizes the creation of monoclonal antibodies against a local strain of the Bc48 gene of B. caballi, a cause of equine piroplasmosis, and the development of a rapid detection test. This test outperformed a comparable commercial ELISA kit test, carrying implications for rapid, efficient diagnosis of B. caballi in horses in the Xinjiang area.
Monoclonal Antibodies Production
- Babesia caballi is a parasite that causes economic harm to the horse industry in China, particularly in the Xinjiang Province, by inducing a disease called equine piroplasmosis.
- Monoclonal antibodies (mAbs) for the Bc48 gene of B. caball were developed, specific to local strains in the Xinjiang area.
- The study describes the production and purification of recombinant Bc48 from Escherichia coli to serve as an immunogen.
- Immunization with the recombinant Bc48 resulted in the production of three distinct hybridoma cells (11F4, 1H2 and 7F4), capable of producing the desired mAbs.
- These antibodies were identified to have a potent reaction with recombinant Bc48 and cells transfected with the Bc48 gene.
Development of the ICT Assay
- An immunochromatographic (ICT) assay was designed using the recombinant Bc48 antigen and the monoclonal antibodies derived in the study.
- The assay was based on colloidal gold labelling, a technique widely used for its simplicity and reliability in rapid detection tests.
- The ICT assay exhibited high sensitivity and specificity for Babesia caballi and no cross-reaction to a similar pathogen, Theileria equi.
Validation of the ICT Assay
- Serum samples from 56 horses in Xinjiang were tested using the newly-developed ICT and a commercial ELISA kit for comparison.
- Results showed that 32 samples were identified as positive by the ICT assay and 33 as positive by the ELISA kit.
- The ICT assay results coincided with the ELISA kit results at a high rate of 98%.
- This result supports the potential of the ICT assay as an efficient diagnostic tool for detecting B. caballi infection in horses specifically in the Xinjiang region.
This study offers hope for improved testing and monitoring of equine piroplasmosis in China, helping to control and mitigate the economic losses associated with the disease in its horse industry.
Cite This Article
APA
Wang P, Song J, Song R, Zhang M, Wu L, Li F, Yan Y, Zhou J, Chahan B, Liao M.
(2019).
Preparation of monoclonal antibodies against Bc48 and development of a rapid detection assay for infection with Babesia caballi in China.
Folia Parasitol (Praha), 66, 2019.005.
https://doi.org/10.14411/fp.2019.005 Publication
Researcher Affiliations
- Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University, Urumqi, Xinjiang, P. R. China.
- Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University, Urumqi, Xinjiang, P. R. China.
- Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University, Urumqi, Xinjiang, P. R. China.
- Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University, Urumqi, Xinjiang, P. R. China.
- Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University, Urumqi, Xinjiang, P. R. China.
- Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University, Urumqi, Xinjiang, P. R. China.
- Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou, P. R. China.
- Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou, P. R. China.
- Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University, Urumqi, Xinjiang, P. R. China.
- Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou, P. R. China.
MeSH Terms
- Animals
- Antibodies, Monoclonal / blood
- Antibodies, Protozoan / blood
- Babesia / immunology
- Babesiosis / diagnosis
- Babesiosis / parasitology
- China
- Female
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Immunoassay / methods
- Immunoassay / veterinary
- Mice
- Mice, Inbred BALB C
- Protozoan Proteins / genetics
- Protozoan Proteins / immunology
- Recombinant Proteins / genetics
- Recombinant Proteins / immunology
- Sensitivity and Specificity
Citations
This article has been cited 2 times.- Srithong P, Chaiyo S, Pasomsub E, Rengpipat S, Chailapakul O, Praphairaksit N. A novel delayed lateral flow immunoassay for enhanced detection of SARS-CoV-2 spike antigen.. Mikrochim Acta 2022 Sep 20;189(10):386.
- Tirosh-Levy S, Gottlieb Y, Fry LM, Knowles DP, Steinman A. Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny.. Pathogens 2020 Nov 8;9(11).
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