FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]2020; 51(3); 1459-1462; doi: 10.1007/s42770-020-00275-4

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.
Get Full Text
Publication Date: 2020-05-03 PubMed ID: 32363568PubMed Central: PMC7455628DOI: 10.1007/s42770-020-00275-4Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Evaluation Study
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

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

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]
ISSN: 1678-4405
NlmUniqueID: 101095924
Country: Brazil
Language: English
Volume: 51
Issue: 3
Pages: 1459-1462

Researcher Affiliations

Ramos, Carolina Pantuzza
  • Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
Lopes, Emily Oliveira
  • Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
Oliveira Júnior, Carlos Augusto
  • Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
Diniz, Amanda Nádia
  • Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
Lobato, Francisco Carlos Faria
  • Veterinary School, Universidade Federal de Minas Gerais, Antônio Carlos Avenue, Belo Horizonte, MG, Brazil.
Silva, Rodrigo Otávio Silveira
  • 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.

References

This article includes 27 references
  1. Czepiel J, Dróżdż M, Pituch H, Kuijper EJ, Perucki W, Mielimonka A, Goldman S, Wultańska D, Garlicki A, Biesiada G. Clostridium difficile infection: review.. Eur J Clin Microbiol Infect Dis 2019 Jul;38(7):1211-1221.
    doi: 10.1007/s10096-019-03539-6pmc: PMC6570665pubmed: 30945014google scholar: lookup
  2. Wilkins TD, Lyerly DM. Clostridium difficile testing: after 20 years, still challenging.. J Clin Microbiol 2003 Feb;41(2):531-4.
    doi: 10.1128/JCM.41.2.531-534.2003pmc: PMC149726pubmed: 12574241google scholar: lookup
  3. Andrés-Lasheras S, Martín-Burriel I, Mainar-Jaime RC, Morales M, Kuijper E, Blanco JL, Chirino-Trejo M, Bolea R. Preliminary studies on isolates of Clostridium difficile from dogs and exotic pets.. BMC Vet Res 2018 Mar 9;14(1):77.
    doi: 10.1186/s12917-018-1402-7pmc: PMC5845233pubmed: 29523201google scholar: lookup
  4. Weese JS. Clostridium (Clostridioides) difficile in animals.. J Vet Diagn Invest 2020 Mar;32(2):213-221.
    pmc: PMC7081495pubmed: 31904312doi: 10.1177/1040638719899081google scholar: lookup
  5. Rodriguez C, Taminiau B, Van Broeck J, Delmée M, Daube G. Clostridium difficile in Food and Animals: A Comprehensive Review.. Adv Exp Med Biol 2016;932:65-92.
    doi: 10.1007/5584_2016_27pubmed: 27350639google scholar: lookup
  6. Silva RO, Ribeiro MG, Palhares MS, Borges AS, Maranhão RP, Silva MX, Lucas TM, Olivo G, Lobato FC. Detection of A/B toxin and isolation of Clostridium difficile and Clostridium perfringens from foals.. Equine Vet J 2013 Nov;45(6):671-5.
    doi: 10.1111/evj.12046pubmed: 23452044google scholar: lookup
  7. Hensgens MP, Keessen EC, Squire MM, Riley TV, Koene MG, de Boer E, Lipman LJ, Kuijper EJ. Clostridium difficile infection in the community: a zoonotic disease?. Clin Microbiol Infect 2012 Jul;18(7):635-45.
    doi: 10.1111/j.1469-0691.2012.03853.xpubmed: 22536816google scholar: lookup
  8. McDonald LC, Gerding DN, Johnson S, Bakken JS, Carroll KC, Coffin SE, Dubberke ER, Garey KW, Gould CV, Kelly C, Loo V, Shaklee Sammons J, Sandora TJ, Wilcox MH. Clinical Practice Guidelines for Clostridium difficile Infection in Adults and Children: 2017 Update by the Infectious Diseases Society of America (IDSA) and Society for Healthcare Epidemiology of America (SHEA).. Clin Infect Dis 2018 Mar 19;66(7):e1-e48.
    doi: 10.1093/cid/cix1085pmc: PMC6018983pubmed: 29462280google scholar: lookup
  9. Crobach MJ, Planche T, Eckert C, Barbut F, Terveer EM, Dekkers OM, Wilcox MH, Kuijper EJ. European Society of Clinical Microbiology and Infectious Diseases: update of the diagnostic guidance document for Clostridium difficile infection.. Clin Microbiol Infect 2016 Aug;22 Suppl 4:S63-81.
    doi: 10.1016/j.cmi.2016.03.010pubmed: 27460910google scholar: lookup
  10. Cançado GGL, Silva ROS, Nader AP, Lobato FCF, Vilela EG. Impact of simultaneous glutamate dehydrogenase and toxin A/B rapid immunoassay on Clostridium difficile diagnosis and treatment in hospitalized patients with antibiotic-associated diarrhea in a university hospital of Brazil.. J Gastroenterol Hepatol 2018 Feb;33(2):393-396.
    doi: 10.1111/jgh.13901pubmed: 28730697google scholar: lookup
  11. Silva RO, Santos RL, Pires PS, Pereira LC, Pereira ST, Duarte MC, de Assis RA, Lobato FC. Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil.. Braz J Microbiol 2013;44(1):133-7.
    doi: 10.1590/S1517-83822013005000008pmc: PMC3804189pubmed: 24159295google scholar: lookup
  12. Silva ROS, Salvarani FM, da Cruz Júnior ECC, Pires PS, Santos RLR, de Assis RA, de Guedes RMC, Lobato FCF. Detection of enterotoxin A and cytotoxin B, and isolation of Clostridium difficile in piglets in Minas Gerais, Brazil.. Ciênc Rural 2011;41:1430–1435.
    doi: 10.1590/S0103-84782011005000100google scholar: lookup
  13. Knight DR, Squire MM, Riley TV. Laboratory detection of Clostridium difficile in piglets in Australia.. J Clin Microbiol 2014 Nov;52(11):3856-62.
    doi: 10.1128/JCM.01225-14pmc: PMC4313215pubmed: 25122859google scholar: lookup
  14. Keessen EC, Hopman NE, van Leengoed LA, van Asten AJ, Hermanus C, Kuijper EJ, Lipman LJ. Evaluation of four different diagnostic tests to detect Clostridium difficile in piglets.. J Clin Microbiol 2011 May;49(5):1816-21.
    doi: 10.1128/JCM.00242-11pmc: PMC3122649pubmed: 21411571google scholar: lookup
  15. Weese JS, Staempfli HR, Prescott JF. A prospective study of the roles of clostridium difficile and enterotoxigenic Clostridium perfringens in equine diarrhoea.. Equine Vet J 2001 Jul;33(4):403-9.
    doi: 10.2746/042516401776249534pubmed: 11469775google scholar: lookup
  16. Anderson MA, Songer JG. Evaluation of two enzyme immunoassays for detection of Clostridium difficile toxins A and B in swine.. Vet Microbiol 2008 Apr 1;128(1-2):204-6.
    doi: 10.1016/j.vetmic.2007.09.022pubmed: 18006252google scholar: lookup
  17. Silva ROS, de Guedes RMC, Silva MX, Lobato FCF. Evaluation of three enzyme immunoassays and toxigenic culture for diagnosis of Clostridium difficile-associated enteritis in piglets.. J Swine Health Prod 2013;21:300–303.
  18. Eastwood K, Else P, Charlett A, Wilcox M. Comparison of nine commercially available Clostridium difficile toxin detection assays, a real-time PCR assay for C. difficile tcdB, and a glutamate dehydrogenase detection assay to cytotoxin testing and cytotoxigenic culture methods.. J Clin Microbiol 2009 Oct;47(10):3211-7.
    doi: 10.1128/JCM.01082-09pmc: PMC2756932pubmed: 19710274google scholar: lookup
  19. Silva RO, Vilela EG, Neves MS, Lobato FC. Evaluation of three enzyme immunoassays and a nucleic acid amplification test for the diagnosis of Clostridium difficile-associated diarrhea at a university hospital in Brazil.. Rev Soc Bras Med Trop 2014 Jul;47(4):447-50.
    doi: 10.1590/0037-8682-0100-2014pubmed: 25229284google scholar: lookup
  20. Silva ROS, Guedes RM de C, Lobato FCF. Clostridium difficile infection: main features and occurrence in domestic species in Brazil.. Ciênc Rural 43:73–80.
    doi: 10.1590/s0103-84782012005000137google scholar: lookup
  21. Medina-Torres CE, Weese JS, Staempfli HR. Validation of a commercial enzyme immunoassay for detection of Clostridium difficile toxins in feces of horses with acute diarrhea.. J Vet Intern Med 2010 May-Jun;24(3):628-32.
    doi: 10.1111/j.1939-1676.2010.00506.xpubmed: 20384955google scholar: lookup
  22. Silva ROS, Neves MS, Ribeiro MG, Palhares MS, de Albuquerque Maranhão RP, Faria Lobato FC. Evaluation of three enzyme immunoassays for diagnosis of Clostridium difficile–associated diarrhea in foals.. J Equine Vet Sci 2014;34:1032–1035.
    doi: 10.1016/j.jevs.2014.05.012google scholar: lookup
  23. Diab SS, Songer G, Uzal FA. Clostridium difficile infection in horses: a review.. Vet Microbiol 2013 Nov 29;167(1-2):42-9.
    doi: 10.1016/j.vetmic.2013.03.032pubmed: 23642413google scholar: lookup
  24. Chapman AM. Acute diarrhea in hospitalized horses.. Vet Clin North Am Equine Pract 2009 Aug;25(2):363-80.
    doi: 10.1016/j.cveq.2009.05.001pubmed: 19580946google scholar: lookup
  25. Diab SS, Rodriguez-Bertos A, Uzal FA. Pathology and diagnostic criteria of Clostridium difficile enteric infection in horses.. Vet Pathol 2013 Nov;50(6):1028-36.
    doi: 10.1177/0300985813489039pubmed: 23686768google scholar: lookup
  26. Clooten J, Kruth S, Arroyo L, Weese JS. Prevalence and risk factors for Clostridium difficile colonization in dogs and cats hospitalized in an intensive care unit.. Vet Microbiol 2008 May 25;129(1-2):209-14.
    doi: 10.1016/j.vetmic.2007.11.013pubmed: 18164560google scholar: lookup
  27. Weese JS, Armstrong J. Outbreak of Clostridium difficile-associated disease in a small animal veterinary teaching hospital.. J Vet Intern Med 2003 Nov-Dec;17(6):813-6.
    doi: 10.1111/j.1939-1676.2003.tb02519pmc: PMC7202293pubmed: 14658717google scholar: lookup

Citations

This article has been cited 5 times.
  1. Luppi A, D'Annunzio G, Torreggiani C, Martelli P. Diagnostic Approach to Enteric Disorders in Pigs. Animals (Basel) 2023 Jan 18;13(3).
    doi: 10.3390/ani13030338pubmed: 36766227google scholar: lookup
  2. Hain-Saunders NMR, Knight DR, Bruce M, Riley TV. Clostridioides difficile infection and One Health: an equine perspective. Environ Microbiol 2022 Mar;24(3):985-997.
    doi: 10.1111/1462-2920.15898pubmed: 35001483google scholar: lookup
  3. Ramos CP, Diniz AN, Leite SM, Lobato FCF, Pereira ST, Rennó MC, de Oliveira Ferreira E, Silva ROS. Evaluation of an immunochromatographic test for the detection of glutamate dehydrogenase for the diagnosis of Clostridioides (Clostridium) difficile infection in dogs. Braz J Microbiol 2021 Dec;52(4):2555-2558.
    doi: 10.1007/s42770-021-00615-ypubmed: 34562233google scholar: lookup
  4. Sedky D, Ghazy AA, Abou-Zeina HAA. Advances in diagnosis of diseases causing diarrhea in newborn calves. Vet Res Commun 2025 Aug 28;49(5):293.
    doi: 10.1007/s11259-025-10855-0pubmed: 40875162google scholar: lookup
  5. Govindaraju R, Govindaraju S, Yun K, Kim J. Fluorescent-Based Neurotransmitter Sensors: Present and Future Perspectives. Biosensors (Basel) 2023 Nov 30;13(12).
    doi: 10.3390/bios13121008pubmed: 38131768google scholar: lookup
Subscribe to our newsletter
Join 99,850 horse owners like you who receive our email updates on horse health and nutrition.