Production and preliminary evaluation of Trypanosoma evansi HSP70 for antibody detection in Equids.
Abstract: The present immuno-diagnostic method using soluble antigens from whole cell lysate antigen for trypanosomosis have certain inherent problems like lack of standardized and reproducible antigens, as well as ethical issues due to in vivo production, that could be alleviated by in vitro production. In the present study we have identified heat shock protein 70 (HSP70) from T. evansi proteome. The nucleotide sequence of T. evansi HSP70 was 2116 bp, which encodes 690 amino acid residues. The phylogenetic analysis of T. evansi HSP70 showed that T. evansi occurred within Trypanosoma clade and is most closely related to T. brucei brucei and T. brucei gambiense, whereas T. congolense HSP70 laid in separate clade. The two partial HSP70 sequences (HSP-1 from N-terminal region and HSP-2 from C-terminal region) were expressed and evaluated as diagnostic antigens using experimentally infected equine serum samples. Both recombinant proteins detected antibody in immunoblot using serum samples from experimental infected donkeys with T. evansi. Recombinant HSP-2 showed comparable antibody response to Whole cell lysate (WCL) antigen in immunoblot and ELISA. The initial results indicated that HSP70 has potential to detect the T. evansi infection and needs further validation on large set of equine serum samples.
Publication Date: 2015-09-27 PubMed ID: 26408598DOI: 10.1515/ap-2015-0104Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study focuses on creating and testing a new method for diagnosing trypanosomosis in equids (horse family), specifically using the heat shock protein 70 (HSP70) from the T. evansi parasite. Initial testing indicates that the proteins captured from HSP70 may be effective in identifying T. evansi infections and reveal it has potential to be further validated with more equine serum samples.
Study Background
- Trypanosomosis is a disease caused by the T. evansi parasite, which affects members of the equid family, such as horses and donkeys.
- The current diagnostic method uses soluble antigens from whole cell lysate antigen, but has issues including lack of standardized, reproducible antigens, and ethical issues due to in vivo (inside a living organism) production.
- This study is aimed at developing a more reliable diagnostic tool using in vitro (outside the organism) production, specifically by identifying and using the heat shock protein 70 (HSP70) from the T. evansi parasite.
Procedure and Main Findings
- Researches worked on the nucleotide sequence of T. evansi HSP70, which was found to be 2116 bp long and coding for 690 amino acid residues.
- Through phylogenetic analysis (study of evolutionary relationships), it was established that T. evansi HSP70 was most closely related to T. brucei brucei and T. brucei gambiense, with T. congolense HSP70 in a separate clade (branch of evolutionary tree).
- To create a diagnostic antigen, two partial HSP70 sequences, one from the N-terminal region (HSP-1) and one from the C-terminal region (HSP-2), were expressed.
- The produced recombinants were then evaluated as diagnostic antigens using serum samples from experimentally infected donkeys.
- Both recombinant proteins were successful in detecting antibodies in immunoblot tests using serum samples from infused donkeys.
- Additionally, the recombinant HSP-2 showed similar antibody responses to whole cell lysate antigens in both immunoblot and ELISA tests, displaying potential for this new diagnostic method.
Conclusion and Further Work
- Preliminary results indicate that HSP70 could be used as a reliable antigen for detecting T. evansi infections.
- However, researchers note that further validation is required on a larger set of equine serum samples to confirm the effectiveness of HSP70 for diagnostic purposes.
Cite This Article
APA
Kumar J, Chaudhury A, Bera BC, Kumar R, Kumar R, Tatu U, Yadav SC.
(2015).
Production and preliminary evaluation of Trypanosoma evansi HSP70 for antibody detection in Equids.
Acta Parasitol, 60(4), 727-734.
https://doi.org/10.1515/ap-2015-0104 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Antibodies, Protozoan / blood
- Antigens, Protozoan / genetics
- Antigens, Protozoan / immunology
- Antigens, Protozoan / isolation & purification
- Equidae
- HSP70 Heat-Shock Proteins / genetics
- HSP70 Heat-Shock Proteins / immunology
- HSP70 Heat-Shock Proteins / isolation & purification
- Immunoassay / methods
- Recombinant Proteins / genetics
- Recombinant Proteins / immunology
- Recombinant Proteins / isolation & purification
- Trypanosoma / genetics
- Trypanosoma / immunology
- Trypanosomiasis / diagnosis
- Trypanosomiasis / veterinary
Citations
This article has been cited 2 times.- Liu YK, Liu GH, Liu L, Wang AB, Cheng TY, Duan DY. Comparative analysis of the anticoagulant activities and immunogenicity of HSC70 and HSC70(TKD) of Haemaphysalis flava. Parasit Vectors 2022 Nov 5;15(1):411.
- Tounkara M, Boulangé A, Thonnus M, Bringaud F, Bélem AMG, Bengaly Z, Thévenon S, Berthier D, Rivière L. Novel protein candidates for serodiagnosis of African animal trypanosomosis: Evaluation of the diagnostic potential of lysophospholipase and glycerol kinase from Trypanosoma brucei. PLoS Negl Trop Dis 2021 Dec;15(12):e0009985.
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