Development and evaluation of recombinant antigen and monoclonal antibody based competition ELISA for the sero- surveillance of surra in animals.
Abstract: Trypanosoma evansi, a haemoflagellated protozoan parasite, is responsible for chronic as well as the acute debilitating disease called surra in a wide range of herbivores and carnivores including domestic and wild animals. Since the parasite is having wide host range, there is a need for diagnostic test which can detect the T. evansi specific antibody in different species of animals for generating sero-surveillance data. In the present study we developed and evaluated competitive enzyme immunoassay using monoclonal antibodies (MAbs) raised against recombinant variable surface glycoprotein (rVSG) of T. evansi. The immunoreactivity of the developed MAbs (IgG3-subtype) was evaluated by immunoblot as well as ELISA and subsequently used in the development and standardization of competitive ELISA (C-ELISA). Further, the serological data generated from the C-ELISA using reference samples constituting true positive or surely infected (35), true negative (45), sero-positive (225) and sero-negative (215) samples and was analyzed statistically. The true positivity/negativity was determined by thin blood smear examination and diagnostic PCR assay, While, seropositivity/seronegativity of the reference samples was determined through standard reference tests. The data showed the diagnostic sensitivity of 92.6% and specificity of 96.4% with Cohen's kappa value of 0.88. In order to determine the utility of C-ELISA in detecting T. evansi antibodies in different species of animals, the assay was further evaluated with 1361 field sera sample comprising bovine, horse, donkey and camel. Since the C-ELISA described herein has showed high sensitivity and specificity, this single test can be explored in the sero-surveillance of T. evansi in a wide range of animals.
Copyright © 2018 Elsevier B.V. All rights reserved.
Publication Date: 2018-07-03 PubMed ID: 30056943DOI: 10.1016/j.jim.2018.06.013Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Animal Health
- Animal Models
- Animal Studies
- Antibodies
- Diagnostic Technique
- Disease Surveillance
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Health
- Immunology
- Infection
- Infectious Disease
- Monoclonal Antibodies
- Protozoa
- Serodiagnosis
- Serological Surveys
- Seroprevalence
- Veterinary Medicine
- Veterinary Research
- Veterinary Science
Summary
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The research article describes the development and evaluation of a diagnostic test (competitive ELISA) utilizing both monoclonal antibodies and a recombinant antigen for the detection of Trypanosoma evansi, a parasite responsible for a disease called surra in many herbivores and carnivores.
Objective and Importance of the Study
- The primary objective of this research was to develop a diagnostic tool that is capable of detecting antibodies specific to T. evansi in a variety of animal species.
- This detection is crucial because T. evansi causes surra, a disease that can impact numerous wild and domestic animals, both herbivores and carnivores, making it a significant concern for animal health and conservation.
Methodology and Evaluation of the Diagnostic Test
- The researchers created this diagnostic test, known as a competitive enzyme immunoassay (C-ELISA), by using monoclonal antibodies (MAbs) produced against the variable surface glycoprotein (VSG) of T. evansi.
- The immunoreactivity of the developed MAbs was checked by immunoblot and ELISA techniques to ensure its success in identifying T. evansi.
- They then used the resultant MAbs in the standardization of the C-ELISA.
- The efficacy of the C-ELISA was assessed using different reference samples including true positive or surely infected, true negative, sero-positive and sero-negative samples.
- The results, showing a diagnostic sensitivity of 92.6% and specificity of 96.4%, were analyzed statistically to ensure their reliability.
Further Testing and Application
- To ensure the assay’s applicability across a variety of animal species, the C-ELISA was tested on field sera samples from bovines, horses, donkeys, and camels.
- Given the high sensitivity and specificity of the C-ELISA, the research suggests that this single test can be employed as a sero-surveillance tool for T. evansi across a wide range of animals, thereby assisting in the prevention and control of the surra disease.
Cite This Article
APA
Sengupta PP, Rudramurthy GR, Ligi M, Jacob SS, Rahman H, Roy P.
(2018).
Development and evaluation of recombinant antigen and monoclonal antibody based competition ELISA for the sero- surveillance of surra in animals.
J Immunol Methods, 460, 87-92.
https://doi.org/10.1016/j.jim.2018.06.013 Publication
Researcher Affiliations
- ICAR-National Institute of Veterinary Epidemiology & Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru 560064, Karnataka, India. Electronic address: pinakiprasad_s@rediffmail.com.
- ICAR-National Institute of Veterinary Epidemiology & Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru 560064, Karnataka, India.
- ICAR-National Institute of Veterinary Epidemiology & Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru 560064, Karnataka, India.
- ICAR-National Institute of Veterinary Epidemiology & Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru 560064, Karnataka, India.
- ICAR-National Institute of Veterinary Epidemiology & Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru 560064, Karnataka, India.
- ICAR-National Institute of Veterinary Epidemiology & Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru 560064, Karnataka, India.
MeSH Terms
- Animals
- Antibodies, Monoclonal, Murine-Derived / chemistry
- Antibodies, Protozoan / chemistry
- Antigens, Protozoan / genetics
- Antigens, Protozoan / immunology
- Buffaloes
- Camelus
- Cattle
- Enzyme-Linked Immunosorbent Assay
- Equidae
- Horses
- Immunoblotting
- Mice
- Mice, Inbred BALB C
- Protozoan Proteins / genetics
- Protozoan Proteins / immunology
- Recombinant Proteins / genetics
- Recombinant Proteins / immunology
- Trypanosoma / genetics
- Trypanosoma / immunology
- Trypanosomiasis / diagnosis
- Trypanosomiasis / immunology
- Trypanosomiasis / veterinary
Citations
This article has been cited 1 times.- Li G, Feng J, Quan K, Sun Z, Yin Y, Yin Y, Chen S, Qin T, Peng D, Liu X. Generation of an avian influenza DIVA vaccine with a H3-peptide replacement located at HA2 against both highly and low pathogenic H7N9 virus. Virulence 2022 Dec;13(1):530-541.
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