Back to Search
Start Over
Comparative evaluation of recombinant HSP70 (N & C-terminal) fragments in the detection of equine trypanosomosis
- Source :
- Veterinary Parasitology. 223:77-87
- Publication Year :
- 2016
- Publisher :
- Elsevier BV, 2016.
-
Abstract
- Trypanosomosis (Surra) is an economically important disease caused by Trypanosoma evansi which is an extracellular parasite present in the plasma, tissues and other body fluids of a wide range of hosts including domesticated animals. Currently, serological reports are based on detection of antibodies by ELISA using whole cell lysate (WCL) antigen, which has a limitation of persistence of anti-trypanosomal antibodies after successful treatment of the disease. Moreover, it has some ethical issues also like requirement of mice for in vivo maintenance of parasite for preparing the antigen. Therefore, in the present study, an attempt was made to evaluate the in vitro production of recombinant heat shock protein 70 (HSP70) for detection of antibodies in experimentally infected ponies. The amino acid sequence analysis of HSP70 revealed that N-terminal region of the protein was highly conserved while the C-terminal region was most divergent. The four different regions of HSP70 protein viz. HSP-1, HSP-2, HSP-3 and HSP-4 were cloned and expressed, among which HSP-1 (N-terminal region) & HSP-2 (C-terminal region) were truncated while HSP-3 & HSP-4 were complete C-terminal proteins. The recombinant fragments were probed with sequentially pooled experimental serum samples where antibodies were detected in these fragments from 10(th) day post infection till the termination of the experiment. Further, these recombinant fragments were also comparatively evaluated with WCL antigen in ELISA using experimental as well as field serum samples. It was observed that after successful treatment of infected ponies, there was a sharp fall in antibodies (within 90 days) when tested with recombinant HSP's fragments, while antibodies persisted even after 469 days when tested against WCL antigen. The sensitivity and specificity of all HSP70 fragments were also estimated from field serum samples with reference to WCL antigen ELISA. The HSP-1 showed minimum sensitivity (41.03%) among all the recombinant fragments. Among the C-terminal fragments, maximum sensitivity was observed with the HSP-2 (61.54%) while minimum was observed with HSP-4 (48.72%). The specificity increases for recombinant fragments from N-terminal to C-terminal region of protein and maximum specificity was observed with HSP-4 fragment (91.3%).
- Subjects :
- 0301 basic medicine
040301 veterinary sciences
Parasitemia
Sensitivity and Specificity
law.invention
Serology
0403 veterinary science
03 medical and health sciences
Antigen
Trypanosomiasis
law
Animals
Parasite hosting
HSP70 Heat-Shock Proteins
Serologic Tests
Amino Acid Sequence
Horses
Cloning, Molecular
Peptide sequence
General Veterinary
biology
Quinolinium Compounds
04 agricultural and veterinary sciences
General Medicine
030108 mycology & parasitology
Trypanosoma evansi
Surra
biology.organism_classification
Virology
Molecular biology
Recombinant DNA
biology.protein
Female
Horse Diseases
Parasitology
Antibody
Subjects
Details
- ISSN :
- 03044017
- Volume :
- 223
- Database :
- OpenAIRE
- Journal :
- Veterinary Parasitology
- Accession number :
- edsair.doi.dedup.....1723987fb060f2636f599af079806528