title: 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.
authors.forename: Jaideep
authors.initials: J
authors.lastname: Kumar
authors.forename: Ashok
authors.initials: A
authors.lastname: Chaudhury
authors.forename: Bidhan C
authors.initials: BC
authors.lastname: Bera
authors.forename: Ritesh
authors.initials: R
authors.lastname: Kumar
authors.forename: Rajender
authors.initials: R
authors.lastname: Kumar
authors.forename: Utpal
authors.initials: U
authors.lastname: Tatu
authors.forename: Suresh Chandra
authors.initials: SC
authors.lastname: Yadav
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D018397:Equidae; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007118:Immunoassay / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014345:Trypanosoma / Q000235:genetics / Q000276:immunology*; D014352:Trypanosomiasis / Q000175:diagnosis / Q000662:veterinary*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins