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Evaluating an indirect rMPSP enzyme-linked immunosorbent assay for the detection of bovine Theileria infection in China

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Abstract

Bovine theileriosis, a tick-borne protozoan disease caused by Theileria annulata, Theileria orientalis and Theileria sinensis, is widespread in China and is a serious economic problem for the Chinese livestock industry. In this study, recombinant major piroplasma surface proteins (MPSP) of T. annulata, T. orientalis and T. sinensis based on MPSP genes were expressed in Escherichia coli BL21(DE3). The immunogenicity and specificity of the three purified recombinant MPSP proteins were evaluated with the reference positive sera of T. annulata, T. orientalis, T. sinensis, Babesia bovis, B abesia bigemina, Babesia major, Babesia motasi, Theileria luwenshuni, Theileria uilenbergi and Anaplasma ovis using an enzyme-linked immunosorbent assay (ELISA) or western blotting. The results showed that all three of the rMPSP proteins had a strong reaction with the sera from cattle infected with T. annulata, T. orientalis and T. sinensis via western blotting but not with other piroplasma and Anaplasma species. Then, the rMPSP protein of T. sinensis was used to develop an iELISA for detecting the three Theileria species infections. The specificity and sensitivity were 95.7 and 95.5 %, respectively, with a threshold of 28.8 % of the specific mean antibody rate (AbR). Finally, 2473 field-collected bovine sera, from 42 prefectures of 17 provinces in China, were tested using the ELISA to evaluate the prevalence of bovine theileriosis, and the average positive rate was 43.6 %. The developed iELISA could be a suitable tool to detect the three bovine Theileria species, and the data also provided important information regarding the current prevalence of bovine theileriosis in China.

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Acknowledgements

This work was financially supported by the 973 Program (2015CB150300); Supporting Program (2013BAD12B03, 2013BAD12B05), MOST, China; NSFC (No. 31402189, No. 31372432, No. 31201899, No. 31272556, No. 31471967), ASTIP, FRIP (2014ZL010) and CAAS; Creative Research Groups of Gansu Province (No. 1210RJIA006); “948” (2014-S05), NBCIS CARS-38, Special Fund for Agro-scientific Research in the Public Research (No. 201303035, No. 201303037), MOA; and Jiangsu Co-innovation Center program for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, State Key Laboratory of Veterinary Etiological Biology Project. The work was also facilitated by CRP No. 16198/R0 IAEA.

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  1. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou, Gansu, 730046, People’s Republic of China

    Shuaiyang Zhao, Junlong Liu, Hongxi Zhao, Youquan Li, Junren Xie, Aihong Liu, Muhammad-Adeel Hassan, Hong Yin, Guiquan Guan & Jianxun Luo

  2. Agricultural College of Ningxia University, Yinchuan, 750021, People’s Republic of China

    Hongxi Zhao

  3. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, People’s Republic of China

    Hong Yin

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  1. Shuaiyang Zhao
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  3. Hongxi Zhao
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Correspondence to Guiquan Guan or Jianxun Luo.

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Zhao, S., Liu, J., Zhao, H. et al. Evaluating an indirect rMPSP enzyme-linked immunosorbent assay for the detection of bovine Theileria infection in China. Parasitol Res 116, 667–676 (2017). https://doi.org/10.1007/s00436-016-5332-7

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