Immunochromatographic test and ELISA for the detection of glutamate dehydrogenase (GDH) and A/B toxins as an alternative for the diagnosis of Clostridioides (Clostridium) difficile-associated diarrhea in foals and neonatal piglets.
Abstract: Considering the lack of studies evaluating the performance of commercially available methods for diagnosis of Clostridioides (Clostridium) difficile infection (CDI) in animals, the present study aimed to assess an immunochromatographic test for detection of glutamate dehydrogenase (GDH) and A/B toxins of C. difficile, also evaluated by an ELISA kit, in foals and neonatal piglets. Intestinal contents of 47 piglets and feces of 35 foals were tested to GDH antigen and A/B toxins in a lateral flow method (Ecodiagnostica, Brazil). Also, these samples were submitted to A/B toxin detection by an ELISA kit (C. difficile Tox A/B II, Techlab Inc., USA), using the toxigenic culture (TC) as the reference method. The GDH component of the lateral flow test showed sensitivity and negative predictive value (NPV) of 100% and a high specificity in samples of piglets (82.61%) and foals (100%). Detection of A/B toxins using the lateral flow test and the ELISA resulted in a specificity of 100% in samples of both species. On the other hand, the sensibility ranged from 54.2 to 90% for the ELISA and from 12.5 to 60% for the lateral flow test for piglets' and foals' samples, respectively. In conclusion, the present work suggests that the lateral flow test for GDH detection could be a useful method for diagnosing CDI in these species. On the other hand, the low sensitivity of the lateral flow test for A/B toxins might compromise its utility in piglets.
Publication Date: 2020-05-03 PubMed ID: 32363568PubMed Central: PMC7455628DOI: 10.1007/s42770-020-00275-4Google Scholar: Lookup
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Summary
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The research is focusing on the development of an immunochromatographic test for the detection of toxins A/B and the glutamate dehydrogenase (GDH) antigen in the diagnoses of Clostridioides (Clostridium) difficile infection in piglets and foals. This method was found to be promising for detecting GDH but less effective for A/B toxins.
Objective of the Research
- The main purpose of this research was to assess and analyze the performance of an immunochromatographic test (lateral flow method) for detecting the GDH and toxins A/B of C. difficile. The study was conducted on both piglets and foals.
- An ELISA kit was also used in parallel for evaluating the presence of A/B toxins. The samples were provided from the feces of 35 foals and the intestinal contents of 47 piglets.
- The performance of the immunochromatographic test and ELISA was compared with the results of the traditional method, a toxigenic culture (TC).
Research Findings
- In the lateral flow test, the GDH component revealed a perfect sensitivity and negative predictive value (NPV) of 100%, indicating a high accuracy in detecting the absence of the antigen. Its specificity was high in samples of both piglets (82.61%) and foals (100%)
- Tests for the detection of toxins A/B using both the lateral flow method and ELISA achieved a specificity of 100%, demonstrating a reliable identification of positive cases in both animal groups.
- However, the sensitivity of these tests varied widely. For ELISA, it ranged from 54.2 to 90%, and for the lateral flow test, it ranged from 12.5 to 60%. These results imply a relatively lower reliability in recognizing negative samples.
Conclusion and Implications
- The lateral flow method for GDH detection, due to its high sensitivity and NPV, was suggested as a useful technique in diagnosing CDI in the examined species.
- However, the low sensitivity of the lateral flow test for A/B toxins detection might limit its practicality, particularly in piglets.
Cite This Article
APA
Ramos CP, Lopes EO, Oliveira Júnior CA, Diniz AN, Lobato FCF, Silva ROS.
(2020).
Immunochromatographic test and ELISA for the detection of glutamate dehydrogenase (GDH) and A/B toxins as an alternative for the diagnosis of Clostridioides (Clostridium) difficile-associated diarrhea in foals and neonatal piglets.
Braz J Microbiol, 51(3), 1459-1462.
https://doi.org/10.1007/s42770-020-00275-4 Publication
Researcher Affiliations
- Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
- Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
- Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
- Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
- Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
- Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil. rodrigo.otaviosilva@gmail.com.
MeSH Terms
- Animals
- Animals, Newborn / microbiology
- Bacterial Proteins / genetics
- Bacterial Proteins / metabolism
- Bacterial Toxins / analysis
- Bacterial Toxins / metabolism
- Brazil
- Clostridioides difficile / enzymology
- Clostridioides difficile / isolation & purification
- Clostridioides difficile / metabolism
- Clostridium Infections / diagnosis
- Clostridium Infections / microbiology
- Clostridium Infections / veterinary
- Diarrhea / diagnosis
- Diarrhea / microbiology
- Diarrhea / veterinary
- Enzyme-Linked Immunosorbent Assay / methods
- Feces / microbiology
- Glutamate Dehydrogenase / analysis
- Glutamate Dehydrogenase / metabolism
- Horse Diseases / diagnosis
- Horse Diseases / microbiology
- Horses
- Immunoassay / methods
- Immunoassay / veterinary
- Swine
- Swine Diseases / diagnosis
- Swine Diseases / microbiology
Grant Funding
- Premio Capes 2015 / Conselho Nacional de Desenvolvimento Cientu00edfico e Tecnolu00f3gico
Conflict of Interest Statement
The authors declare that they have no conflicts of interest.
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Citations
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