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

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.

Development of an Indirect ELISA Based on a Recombinant Chimeric Protein for the Detection of Antibodies against Bovine Babesiosis

“…ROC analysis was applied to the rMABbo-iELISA to assess the cutoff value using a batch of samples of known serological status. The cutoff value of 35% obtained in our study is similar to the value obtained by other studies using iELISA based on recombinant antigens and bovine sera [ 16 , 17 ]. Sensitivity and specificity of the rMABbo-iELISA was optimal and very similar to the values reported for the MZ-iELISA [ 15 ].…”

First Molecular Detection of Piroplasm Infection in Pet Dogs from Gansu, China

“…These parasites are usually found in horses, sheep and cattle. T. sinensis is distributed only in Gansu Province China and is transmitted by Haemaphysalis qinghaiensis (Zhao et al, 2017). In our study, records from dogs infected with B. canis vogeli and T. sinensis provided by veterinarians from clinics indicated that these pet dogs spent most of their time indoors and they had limited exposure to the outside environment with active R. sanguineus and H. qinghaiensis ticks.…”

Oriental Theileriosis

“…It is unsurprising therefore that seroconversion to the MPSP is also strongly correlated with both parasite load and the Ikeda genotype [133]. Another study demonstrated that humoral responses to experimental infection with T. orientalis (via mechanical transfer) are variable and only established after persistent infection [152]. The role of humoral immunity in protecting against T. orientalis infection in adult cattle is unclear.…”

“…While a number of serological tests exist for the detection of T. orientalis [29,87,133,152,162], these assays are currently not genotype-specific and in some cases also cross-react with other Theileria species [152,162]. Serological tests are of a similar sensitivity to blood smear examination and are most reliable when the animals are clinically affected, but are unsuitable for testing newly infected animals that have not yet seroconverted [133].…”