Enzyme-Linked Immunosorbent Assays (ELISA) - strategies used in diagnosis of animal infectious disease for animal health

Enzyme-linked immunosorbent assays (ELISAs) incorporate the sensitivity of simple enzyme assays with the specificity of antibodies, by employing antigens or antibodies coupled to an easily-assayed enzyme. As such ELISA is much more rapid method than immunoblotting to detect specific viral protein from a cell, tissue, organ, or body fluid. There are two main variations of ELISAs: antigen-capture ELISA (detecting viral proteins), involve attachment of a capture antibody to a solid matrix for the viral protein of interest, while antibody-capture ELISA measures the specific antibody level in a sample, by coating viral antigen protein on a solid surface.

There are two principles based on antigen-capture and antibody-capture ELISAs. In a general, ELISAs are considered a highly sensitive method that can detect a fairly low number of proteins at the range of picomolar to nanomolar range (10-12 to 10-9 moles per liter). ELISA method was found useful as a diagnostic tool to detect influenza viral antigen much quicker than other conventional virus detection methods [7]. In another previous study, comparison of ELISA, with conventional methods has demonstrated ELISA superiority for the rapid detection and identification of influenza A virus [8]. A simplified and standardized neutralization enzyme immunoassay (Nt-EIA) was developed to detect measles virus growth in Vero cells and to quantify measles neutralizing antibody [9].

Newer EIA formats for hepatitis C virus diagnostics have been constantly evaluated [10, 11]. As such ELISAs are being used for plethora of application both in experimental and diagnostic virology including dengue, and influenza [12-14]. On the other hand, although rapid than traditional plaque assays or TCID50, ELISA assays sometimes could be quite expensive, due to the cost of reagents used. Unfortunately, sometimes required antibodies may not be commercially developed as well. In contrast, attempts to develop antibodies in-house may be quite expensive. Additional variability may also be introduced due to high background signals generated by non-specific binding, or cross-reactivity with non-viral protein targets.

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Figure 2. A schematic representation of two principles based on antigen or antibody capture ELISA [15].



Category

Genemedi provides diagnostic antibodies and antigens for the in vitro diagnosis of diseases from the Companion Animal, Cat/Feline, Dog/Canine, Rabbit, Bovines/Cattle, Ovines/Sheep, Caprine/Goat, Equine/Horse, Swine/Porcine/Pig, Avian/bird/poultry, Fish and so on.

Animal category

Disease category

Veterinary NCDs (noncommunicable diseases)


  • · Acute inflammation, infections
    · Inflammatory diseases, infections
    · Myocardial injury, heart disease

    · Bacterial infections, sepsis
    · Heart failure, cardiac disease
    · Thrombosis, disseminated intravascular coagulation

Veterinary Infectious Disease


  • · Infectious bursal/Gumboro disease
    · Foot and mouth disease
    · Hemolytic uremic syndrome/E. coli infection

    · Bovine spongiform encephalopathy (BSE)
    · Brucella abortus, Brucella melitensis, Brucellosis
    · Monkeypox, cowpox



Technical resource


The Knowledge base of Diagnostics
  • Immuno-Diagnostics Antibodies & Antigens for Veterinary NCDs (noncommunicable diseases) Test
  • What is animal infectious disease?
  • What is lateral flow and how to use it for animal infectious disease?
  • Enzyme-Linked Immunosorbent Assays (ELISA)
  • Immunofluorescence staining (IF)- strategies for animal health
  • Immunohistochemistry (IHC), In situ hybridization (ISH), Virus isolation (VI)
  • Electron microscopy (EM), Molecular Methods
  • The strategies used in diagnosis of animal infectious disease for animal health

  • Reference:

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    3. Leslie MJ, M.J. Surveillance for zoonotic diseases. BLUKO97- Mikanatha 2007; Available from: http://courses.washington.edu/zepi526/Papers08/Rabies%20chapter.pdf.
    4. Available from: https://www.doctorsgate.com/en/what-are-the-current-diagnostic-tests-for-covid-19/.
    5. Anylab. Available from: www.zetbio.com.
    6. Li, H., et al., A new and rapid approach for detecting COVID-19 based on S1 protein fragments. Clin Transl Med, 2020. 10(2): p. e90.
    7. Khanna, M., et al., Evaluation of influenza virus detection by direct enzyme immunoassay (EIA) and conventional methods in asthmatic patients. J Commun Dis, 2001. 33(3): p. 163-9.
    8. Waner, J.L., et al., Comparison of Directigen FLU-A with viral isolation and direct immunofluorescence for the rapid detection and identification of influenza A virus. J Clin Microbiol, 1991. 29(3): p. 479-82.
    9. Cameron, J.D., A.P. Skubitz, and L.T. Furcht, Type IV collagen and corneal epithelial adhesion and migration. Effects of type IV collagen fragments and synthetic peptides on rabbit corneal epithelial cell adhesion and migration in vitro. Invest Ophthalmol Vis Sci, 1991. 32(10): p. 2766-73.
    10. Kim, M.H., S.Y. Kang, and W.I. Lee, Evaluation of a new rapid test kit to detect hepatitis C virus infection. J Virol Methods, 2013. 193(2): p. 379-82.
    11. Niu, X., et al., [Establishment of the evaluation reference system for domestic anti-hepatitis C virus diagnostic enzyme immunoassay kits]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2013. 29(7): p. 761-4.
    12. Filice, G., et al., Sensitivity and specificity of anti-HIV ELISA employing recombinant (p24, p66, gp120) and synthetic (gp41) viral antigenic peptides. Microbiologica, 1991. 14(3): p. 185-94.
    13. de Boer, G.F., W. Back, and A.D. Osterhaus, An ELISA for detection of antibodies against influenza A nucleoprotein in humans and various animal species. Arch Virol, 1990. 115(1-2): p. 47-61.
    14. Cuzzubbo, A.J., et al., Comparison of PanBio dengue duo enzyme-linked immunosorbent assay (ELISA) and MRL dengue fever virus immunoglobulin M capture ELISA for diagnosis of dengue virus infections in Southeast Asia. Clin Diagn Lab Immunol, 1999. 6(5): p. 705-12.
    15. Maria-C-Jimenez-Martinez. Available from:
    https://www.researchgate.net/profile/Maria-C-Jimenez-Martinez/publication/320265684/figure/fig3/AS:547016444395520@1507430291554/ELISA-assays-Direct-ELISA-mostly-used-for-antigen-detection-Indirect-ELISA-mainly-used.png.
    16. sciencephoto. Available from: https://www.sciencephoto.com/media/90184/view.
    17. Uyeki, T.M., et al., Clinical Practice Guidelines by the Infectious Diseases Society of America: 2018 Update on Diagnosis, Treatment, Chemoprophylaxis, and Institutional Outbreak Management of Seasonal Influenzaa. Clin Infect Dis, 2019. 68(6): p. e1-e47.
    18. Goldsmith, C. Hantavirus Life Cycle and Infection Process. Available from: https://www.hantasite.com/2017/03/hantavirus-life-cycle-and-infection.html.
    19. METHODS, M. 2013; 3:207
    20. biotech, G.; Available from: https://www.globalbiotechinsights.com/articles/20247/the-worldwide-test-for-covid-19.




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