Development of a microsphere-based immunoassay for the serological detection of glanders in equids.
Abstract: Burkholderia mallei is the etiologic agent of glanders, an infectious disease of solipeds, with renewed scientific interest due to its increasing incidence in different parts of the world. More rapid, sensitive and specific assays are required by laboratories for confirmatory testing of this disease. A microsphere-based immunoassay consisting of beads coated with B. mallei recombinant proteins (BimA, GroEL, Hcp1, and TssB) has been developed for the serological diagnosis of glanders. The proteins' performance was compared with the OIE reference complement fixation test (CFT) and an indirect enzyme-linked immunosorbent assay (iELISA) on a large panel of sera comprised of uninfected horses (n=198) and clinically confirmed cases of glanders from India and Pakistan (n=99). Using Receiver Operating Characteristics (ROC) analysis and adjusting the cutoff levels, Hcp1 (Se=100%, Sp=99.5%) and GroEL (Se= 97%, Sp=99.5%) antigens exhibited the best specificity and sensitivity. Neither Hcp1 and GroEL proteins, nor iELISA reacted with doubtful and positive CFT samples from glanders free countries which further confirmed the false positive reactions seen in CFT.
Copyright © 2020 Elsevier B.V. All rights reserved.
Publication Date: 2020-04-14 PubMed ID: 32302692DOI: 10.1016/j.actatropica.2020.105463Google Scholar: Lookup
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- Journal Article
Summary
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The research article discusses the development of a more rapid, sensitive and specific microsphere-based immunoassay for the diagnosis of an infectious disease, glanders, in equids (such as horses, donkeys and mules).
Introduction
- The focus of the study is glanders, an infectious disease that primarily affects equids, caused by the Burkholderia mallei bacterium. This disease has gained increased scientific interest due to its rising incidence across the world.
- Existing test methods were found insufficient due to their sensitivity and specificity rates, leading to the need for improved diagnostic assays.
Methodology: Development of a Microsphere-based Immunoassay
- The researchers developed a microsphere-based immunoassay, using beads coated with B. mallei recombinant proteins (BimA, GroEL, Hcp1, and TssB) for the serological diagnosis of glanders.
- This method was tested on a large panel of uninfected equids (198 in total) as well as clinically confirmed glanders cases from India and Pakistan (99 in total).
Result Comparison with Existing Test Methods
- The performance of the newly developed immunoassay proteins was compared with the results achieved through the OIE reference complement fixation test (CFT) and an indirect enzyme-linked immunosorbent assay (iELISA).
- Using Receiver Operating Characteristics (ROC) analysis and after adjusting the cutoff levels, the antigens from two proteins, GroEL and Hcp1, showed the best performance with a sensitivity (Se) of 100% and 97% and specificity (Sp) of 99.5% for both.
Conclusion
- The novel microsphere-based immunoassay has been found to be more specific and sensitive compared to the previously used CFT and iELISA methods.
- Both proteins GroEL and Hcp1 as well as the iELISA did not react to doubtful and positive CFT samples from countries free of glanders. This indicates that they avoid the false positives often encountered with the CFT method, further confirming their superior performance in diagnosing glanders.
Cite This Article
APA
Laroucau K, Saqib M, Martin B, Deshayes T, Bertin C, Wernery U, Joseph S, Singha H, Tripathi BN, Beck C.
(2020).
Development of a microsphere-based immunoassay for the serological detection of glanders in equids.
Acta Trop, 207, 105463.
https://doi.org/10.1016/j.actatropica.2020.105463 Publication
Researcher Affiliations
- Paris-Est University, ANSES, Animal Health Laboratory, Bacterial Zoonosis Unit, EURL for equine diseases/Glanders, Maisons-Alfort, France. Electronic address: karine.laroucau@anses.fr.
- Department of Clinical Medicine and Surgery, University of agriculture, Faisalabad, Pakistan.
- Paris-Est University, ANSES, Animal Health Laboratory, Bacterial Zoonosis Unit, EURL for equine diseases/Glanders, Maisons-Alfort, France.
- Paris-Est University, ANSES, Animal Health Laboratory, Bacterial Zoonosis Unit, EURL for equine diseases/Glanders, Maisons-Alfort, France.
- Paris-Est University, ANSES, Animal Health Laboratory, Bacterial Zoonosis Unit, EURL for equine diseases/Glanders, Maisons-Alfort, France.
- Central Veterinary Research Laboratory, PO Box 597, Dubai, United Arab Emirates.
- Central Veterinary Research Laboratory, PO Box 597, Dubai, United Arab Emirates.
- ICAR-National Research Centre on Equines, Hisar, India.
- ICAR-National Research Centre on Equines, Hisar, India.
- UMR 1161 Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EURL for equine diseases, Maisons-Alfort, France.
MeSH Terms
- Animals
- Burkholderia mallei / immunology
- Complement Fixation Tests
- Enzyme-Linked Immunosorbent Assay
- Glanders / diagnosis
- Horses
- Microspheres
- Serologic Tests
Conflict of Interest Statement
Declaration of Competing Interest None of the authors have any financial or personal relationships with any individuals or organisations that could inappropriately influence or bias this paper.
Citations
This article has been cited 7 times.- Ichikawa Y, Iinuma Y, Okagawa T, Shimbo R, Enkhtuul B, Khurtsbaatar O, Kinoshita Y, Niwa H, Aoshima K, Kobayashi A, Batbaatar V, Ohashi K, Kimura T. Comparison of immunogenicity of 17 Burkholderia mallei antigens and whole cell lysate using indirect ELISA. J Vet Med Sci 2025 Apr 1;87(4):394-401.
- Gasqué M, Guernier-Cambert V, Manuel G, Aaziz R, Terret J, Deshayes T, Baudrimont X, Breurec S, Rochelle-Newall E, Laroucau K. Reassessing the distribution of Burkholderia pseudomallei outside known endemic areas using animal serological screening combined with environmental surveys: The case of Les Saintes (Guadeloupe) and French Guiana. PLoS Negl Trop Dis 2024 Sep;18(9):e0011977.
- Desoutter A, Deshayes T, Vorimore F, Klotoe B, Durand B, Colot J, Wagner-Lichtenegger G, Steinmetz I, Tuanyok A, Laroucau K. Isolation of Burkholderia pseudomallei from a goat in New Caledonia: implications for animal and human health monitoring and serological tool comparison. BMC Vet Res 2024 Mar 23;20(1):114.
- Wagner GE, Berner A, Lipp M, Kohler C, Assig K, Lichtenegger S, Saqib M, Müller E, Trinh TT, Gad AM, Söffing HH, Ehricht R, Laroucau K, Steinmetz I. Protein Microarray-Guided Development of a Highly Sensitive and Specific Dipstick Assay for Glanders Serodiagnostics. J Clin Microbiol 2023 Jan 26;61(1):e0123422.
- P K, M M, S M, Kr P, T D, G W, Ma V, K L. First glanders cases detected in Nepal underscore the need for surveillance and border controls. BMC Vet Res 2022 Apr 6;18(1):132.
- Verney M, Gautron M, Lemans C, Rincé A, Hans A, Hébert L. Development of a microsphere-based immunoassay for the serological diagnosis of equine trypanosomosis. Sci Rep 2022 Jan 25;12(1):1308.
- Elschner MC, Melzer F, Singha H, Muhammad S, Gardner I, Neubauer H. Validation of a Commercial Glanders ELISA as an Alternative to the CFT in International Trade of Equidae. Front Vet Sci 2021;8:628389.
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