Frontiers in veterinary science2021; 8; 628389; doi: 10.3389/fvets.2021.628389

Validation of a Commercial Glanders ELISA as an Alternative to the CFT in International Trade of Equidae.

Abstract: Glanders, caused by (.) is a notifiable zoonotic disease in equidae. For international trade and movement of equids, certificates of negative serological test results for antibodies against are required. To date, the complement fixation test (CFT) is the mandatory test to issue these health certificates. The CFT is difficult to standardize and, due to its poor specificity, often leads to false-positive reactions resulting in trade restrictions with considerable financial consequences. In the present study, the new ID Screen Glanders Double Antigen Multispecies ELISA (GLANDA- ELISA) (IDvet, Grabels, France) was evaluated using 400 negative and 370 glanders positive field samples of equidae. The GLANDA-ELISA was significantly more specific (99.8%) than the CFT (97.0%). Considering the comparable sensitivities of CFT (96.5%) and ELISA (98.1%), this new GLANDA-ELISA test appears a suitable confirmatory test and a realistic alternative for serological testing of horses for trade or movement.
Publication Date: 2021-02-16 PubMed ID: 33665218PubMed Central: PMC7920956DOI: 10.3389/fvets.2021.628389Google 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.
  • 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 study focuses on validating a commercial glanders ELISA test as an effective alternative to the traditional CFT in controlling the international trade of equids. The new ELISA test exhibits higher specificity than the CFT, potentially reducing false-positives and associated financial consequences.

Objective of the Research

  • The primary aim of this research is to evaluate and validate the use of a new commercial glanders ELISA (GLANDA-ELISA) developed by IDvet, over the traditional CFT for the international trade and movement of equids.
  • The researchers endeavor to test the ELISA’s accuracy and effectiveness, focusing on its specificity and sensitivity compared to the CFT.

About Glanders and its Diagnostic Issues

  • Glanders is a contagious zoonotic disease affecting equidae caused by the bacterium Burkholderia mallei. It is notifiable, meaning any instance of this disease has to be reported to veterinary authorities.
  • For the international trade and movement of equids, negative results for antibodies against B. mallei are required. Currently, these results are obtained through the complement fixation test (CFT).
  • However, the CFT test has been found to be difficult to standardize and often leads to poor specificity, which leads to false-positive reactions. The false positives result in trade restrictions leading to financial losses.

Evaluation of the GLANDA-ELISA Test

  • The validation process of the GLANDA-ELISA involved testing 400 negative and 370 glanders positive field samples of equidae.
  • Results showed that the new ELISA test was significantly more specific (99.8%) than the CFT (97.0%). This high specificity can reduce the number of false positive reactions, thereby decreasing unnecessary trade restrictions and associated financial implications.

Sensitivity and Suitability of GLANDA-ELISA

  • The research also reported comparable sensitivities for the CFT (96.5%) and the ELISA (98.1%), i.e., they both effectively identified true positive samples.
  • This equivalency in sensitivity, combined with ELISA’s significantly better specificity, makes the GLANDA-ELISA a viable alternative for serological testing of horses for trade or movement.

Cite This Article

APA
Elschner MC, Melzer F, Singha H, Muhammad S, Gardner I, Neubauer H. (2021). Validation of a Commercial Glanders ELISA as an Alternative to the CFT in International Trade of Equidae. Front Vet Sci, 8, 628389. https://doi.org/10.3389/fvets.2021.628389

Publication

ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 8
Pages: 628389
PII: 628389

Researcher Affiliations

Elschner, Mandy Carolina
  • Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Jena, Germany.
Melzer, Falk
  • Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Jena, Germany.
Singha, Harisankar
  • Indian Council of Agricultural Research, National Research Centre on Equines, Hisar, India.
Muhammad, Saqib
  • Department of Clinical Medicine and Surgery, University of Agriculture, Faisalabad, Pakistan.
Gardner, Ian
  • Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island (UPEI), Charlottetown, PE, Canada.
Neubauer, Heinrich
  • Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Jena, Germany.

Conflict of Interest Statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

References

This article includes 27 references
  1. Ghori MT, Khan MS, Khan JA, Rabbani M, Shabbir MZ, Chaudhry HR, Ali MA, Muhammad J, Elschner MC, Jayarao BM. Seroprevalence and risk factors of glanders in working equines - Findings of a cross-sectional study in Punjab province of Pakistan.. Acta Trop 2017 Dec;176:134-139.
  2. Arun S, Neubauer H, Gu00fcrel A, Ayyildiz G, Kusu00e7u B, Yesildere T, Meyer H, Hermanns W. Equine glanders in Turkey.. Vet Rec 1999 Mar 6;144(10):255-8.
    doi: 10.1136/vr.144.10.255pubmed: 10209817google scholar: lookup
  3. Mota RA, da Fonseca Oliveira AA, da Silva AM, Junior JW, da Silva LB, de Farias Brito M, Rabelo SS. Glanders in donkeys (Equus Asinus) in the state of pernambuco, Brazil: A case report.. Braz J Microbiol 2010 Jan;41(1):146-9.
  4. Malik P, Singha H, Goyal SK, Khurana SK, Tripathi BN, Dutt A, Singh D, Sharma N, Jain S. Incidence of Burkholderia mallei infection among indigenous equines in India.. Vet Rec Open 2015;2(2):e000129.
    doi: 10.1136/vetreco-2015-000129pmc: PMC4594314pubmed: 26457190google scholar: lookup
  5. Singha H, Shanmugasundaram K, Tripathi BN, Saini S, Khurana SK, Kanani A, Shah N, Mital A, Kanwar P, Bhatt L, Limaye V, Khasa V, Arora R, Gupta S, Sangha S, Sharma H, Agarwal SK, Tapase J, Parnam S, Dubey P, Baalasundaram SK, Mandal BN, Virmani N, Gulati BR, Malik P. Serological surveillance and clinical investigation of glanders among indigenous equines in India from 2015 to 2018.. Transbound Emerg Dis 2020 May;67(3):1336-1348.
    doi: 10.1111/tbed.13475pubmed: 31916415google scholar: lookup
  6. Erdemsurakh O, Ochirbat K, Gombosuren U, Tserendorj B, Purevdorj B, Vanaabaatar B, Aoshima K, Kobayashi A, Kimura T. Seroprevalence of equine glanders in horses in the central and eastern parts of Mongolia.. J Vet Med Sci 2020 Sep 24;82(9):1247-1252.
    doi: 10.1292/jvms.20-0219pmc: PMC7538334pubmed: 32641602google scholar: lookup
  7. Scholz HC, Pearson T, Hornstra H, Projahn M, Terzioglu R, Wernery R, Georgi E, Riehm JM, Wagner DM, Keim PS, Joseph M, Johnson B, Kinne J, Jose S, Hepp CM, Witte A, Wernery U. Genotyping of Burkholderia mallei from an outbreak of glanders in Bahrain suggests multiple introduction events.. PLoS Negl Trop Dis 2014 Sep;8(9):e3195.
  8. OIE . Chapter 2.5.11: Glanders and melioidosis. In: Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. (2018). Available online at: http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.05.11_GLANDERS.pdf (accessed January 25, 2021).
  9. de Carvalho Filho MB, Ramos RM, Fonseca AA Jr, de Lima Orzil L, Sales ML, de Assis Santana VL, de Souza MM, Dos Reis Machado E, Filho PR, Leite RC, Dos Reis JK. Development and validation of a method for purification of mallein for the diagnosis of glanders in equines.. BMC Vet Res 2012 Sep 2;8:154.
    doi: 10.1186/1746-6148-8-154pmc: PMC3514244pubmed: 22937975google scholar: lookup
  10. da Silva KP, de Campos Takaki GM, da Silva LB, Saukas TN, Santos AS, Mota RA. Assessment of the effectiveness of the PPD-mallein produced in Brazil for diagnosing glanders in mules.. Braz J Microbiol 2013;44(1):179-81.
  11. Adhikari N, Acharya KP, Wilson RT. The potential for an outbreak of glanders in Nepal.. Trop Med Health 2019;47:57.
    doi: 10.1186/s41182-019-0185-2pmc: PMC6905013pubmed: 31889885google scholar: lookup
  12. Neubauer H, Sprague LD, Zacharia R, Tomaso H, Al Dahouk S, Wernery R, Wernery U, Scholz HC. Serodiagnosis of Burkholderia mallei infections in horses: state-of-the-art and perspectives.. J Vet Med B Infect Dis Vet Public Health 2005 Jun;52(5):201-5.
  13. Elschner MC, Scholz HC, Melzer F, Saqib M, Marten P, Rassbach A, Dietzsch M, Schmoock G, de Assis Santana VL, de Souza MM, Wernery R, Wernery U, Neubauer H. Use of a Western blot technique for the serodiagnosis of glanders.. BMC Vet Res 2011 Jan 19;7:4.
    doi: 10.1186/1746-6148-7-4pmc: PMC3034690pubmed: 21247488google scholar: lookup
  14. Elschner MC, Laroucau K, Singha H, Tripathi BN, Saqib M, Gardner I, Saini S, Kumar S, El-Adawy H, Melzer F, Khan I, Malik P, Sauter-Louis C, Neubauer H. Evaluation of the comparative accuracy of the complement fixation test, Western blot and five enzyme-linked immunosorbent assays for serodiagnosis of glanders.. PLoS One 2019;14(4):e0214963.
  15. Khan I, Wieler LH, Melzer F, Gwida M, Santana VL, de Souza MM, Saqib M, Elschner MC, Neubauer H. Comparative evaluation of three commercially available complement fixation test antigens for the diagnosis of glanders.. Vet Rec 2011 Nov 5;169(19):495.
    doi: 10.1136/vr.d5410pubmed: 21896565google scholar: lookup
  16. Khan I, Wieler LH, Saqib M, Melzer F, Santana VL, Neubauer H, Elschner MC. Effect of incubation temperature on the diagnostic sensitivity of the glanders complement fixation test.. Rev Sci Tech 2014 Dec;33(3):869-75.
    doi: 10.20506/rst.33.3.2324pubmed: 25812210google scholar: lookup
  17. Laroucau K, Colaneri C, Jau00ff M, Corde Y, Drapeau A, Durand B, Zientara S, Beck C. Interlaboratory ring trial to evaluate CFT proficiency of European laboratories for diagnosis of glanders in equids.. Vet Rec 2016 Jun 18;178(25):632.
    doi: 10.1136/vr.103617pubmed: 27122499google scholar: lookup
  18. Abreu DC, Gomes AS, Tessler DK, Chiebao DP, Fava CD, Romaldini AHCN, Araujo MC, Pompei J, Marques GF, Harakava R, Pituco EM, Nassar AFC. Systematic monitoring of glanders-infected horses by complement fixation test, bacterial isolation, and PCR.. Vet Anim Sci 2020 Dec;10:100147.
    doi: 10.1016/j.vas.2020.100147pmc: PMC7566944pubmed: 33089006google scholar: lookup
  19. Laroucau K, Saqib M, Martin B, Deshayes T, Bertin C, Wernery U, Joseph S, Singha H, Tripathi BN, Beck C. Development of a microsphere-based immunoassay for the serological detection of glanders in equids.. Acta Trop 2020 Jul;207:105463.
  20. Singha H, Malik P, Goyal SK, Khurana SK, Mukhopadhyay C, Eshwara VK, Singh RK. Optimization and validation of indirect ELISA using truncated TssB protein for the serodiagnosis of glanders amongst equines.. ScientificWorldJournal 2014;2014:469407.
    doi: 10.1155/2014/469407pmc: PMC3932216pubmed: 24672321google scholar: lookup
  21. Khan I, Elschner MC, Melzer F, Gwida M, Wieler LH, Ali R, Saqib M, Neubauer H. Performance of complement fixation test and confirmatory immunoblot as two-cascade testing approach for serodiagnosis of glanders in an endemic region of South East Asia.. Berl Munch Tierarztl Wochenschr 2012 Mar-Apr;125(3-4):117-21.
    pubmed: 22515029
  22. Dohoo I MW, Stryhn H. Veterinary Epidemiologic Research. Charlottetown, PE: VER Inc. (2009).
  23. Povitzky OR. Prompt macroscopic agglutination in the diagnosis of glanders. J Immunol. (1918) 3:463u201379.
  24. Yazdansetad S, Mosavari N, Tadayon K, Mehregan I. Development of an immunoblotting assay for serodiagnosis of Burkholderia mallei infection: the whole-cell proteome-based paradigm.. Iran J Microbiol 2019 Jun;11(3):232-238.
    doi: 10.18502/ijm.v11i3.1326pmc: PMC6711873pubmed: 31523407google scholar: lookup
  25. Singh S, Dohre SK, Kamthan A, Pal V, Karothia BS, Singha HS, Kumar S. Improvement of recombinant-truncated Burkholderia motility protein A (BimA)-based indirect ELISA for equine glanders.. J Immunoassay Immunochem 2018;39(5):565-575.
    doi: 10.1080/15321819.2018.1523800pubmed: 30303469google scholar: lookup
  26. Pal V, Kumar S, Malik P, Rai GP. Evaluation of recombinant proteins of Burkholderia mallei for serodiagnosis of glanders.. Clin Vaccine Immunol 2012 Aug;19(8):1193-8.
    doi: 10.1128/CVI.00137-12pmc: PMC3416092pubmed: 22695165google scholar: lookup
  27. Jacobson RH. Validation of serological assays for diagnosis of infectious diseases.. Rev Sci Tech 1998 Aug;17(2):469-526.
    doi: 10.20506/rst.17.2.1119pubmed: 9713892google scholar: lookup

Citations

This article has been cited 3 times.
  1. Wagner GE, Berner A, Lipp M, Kohler C, Assig K, Lichtenegger S, Saqib M, Mu00fcller E, Trinh TT, Gad AM, Su00f6ffing 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.
    doi: 10.1128/jcm.01234-22pubmed: 36541753google scholar: lookup
  2. 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.
    doi: 10.1186/s12917-022-03233-4pubmed: 35387664google scholar: lookup
  3. Nielsen SS, Alvarez J, Bicout DJ, Calistri P, Canali E, Drewe JA, Garin-Bastuji B, Gonzales Rojas JL, Schmidt CG, Herskin M, Michel V, Miranda Chueca Mu00c1, Padalino B, Pasquali P, Spoolder H, Stu00e5hl K, Velarde A, Viltrop A, Winckler C, Gubbins S, Laroucau K, Antoniou SE, Aznar I, Broglia A, Lima E, Van der Stede Y, Zancanaro G, Roberts HC. Assessment of the control measures of the category A diseases of Animal Health Law: Burkholderia mallei (Glanders).. EFSA J 2022 Jan;20(1):e07069.
    doi: 10.2903/j.efsa.2022.7069pubmed: 35035583google scholar: lookup