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Home / Equine Research Bank / Microorganisms / Serological Hendra Virus Diagnostics Using an Indirect ELISA...
Microorganisms2022; 10(6); 1095; doi: 10.3390/microorganisms10061095

Serological Hendra Virus Diagnostics Using an Indirect ELISA-Based DIVA Approach with Recombinant Hendra G and N Proteins.

Abstract: Since the identification of Hendra virus (HeV) infections in horses in Australia in 1994, more than 80 outbreaks in horses have been reported, and four out of seven spillover infections in humans had a fatal outcome. With the availability of a subunit vaccine based on the HeV-Glycoprotein (HeV-G), there is a need to serologically ifferentiate the nfected from the accinated nimals (DIVA). We developed an indirect ELISA using HeV-G expressed in and HeV-Nucleoprotein (HeV-N) expressed in recombinant baculovirus-infected insect cells as antigens. During evaluation, we tested panels of sera from naïve, vaccinated and infected horses that either originated from a Hendra-virus free region, or had been pre-tested in validated diagnostic tests. Our data confirm the reliability of this approach, as HeV-N-specific antibodies were only detected in sera from infected horses, while HeV-G-specific antibodies were detected in infected and vaccinated horses with a high level of specificity and sensitivity. Given the excellent correlation of data obtained for German and Australian HeV-negative horses, we assume that this test can be applied for the testing of horse serum samples from a variety of geographical regions.
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Publication Date: 2022-05-25 PubMed ID: 35744614PubMed Central: PMC9230382DOI: 10.3390/microorganisms10061095Google Scholar: Lookup
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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 study presents a reliable approach to distinguish between vaccinated and infected horses for Hendra Virus using an Indirect ELISA technique and recombinant Hendra G and N proteins.

Background

  • Hendra virus (HeV) is a severe infection in horses that was first recognized in Australia in 1994, with over 80 outbreaks reported since its identification. It has caused fatalities in humans who were infected due to contact with the infected horses.
  • A subunit vaccine based on HeV-Glycoprotein (HeV-G) has been developed to control the spread of the virus in horses. This created a necessity to differentiate serologically between vaccinated and infected horses.

Objective

  • The objective of this research was to develop an indirect Enzyme-Linked Immunosorbent Assay (ELISA) using HeV-G and HeV-Nucleoprotein (HeV-N) as antigens, which could distinguish the infected from the vaccinated horses.

Methodology

  • The researchers expressed the HeV-G in Escherichia coli (E. coli) and expressed HeV-N in recombinant baculovirus-infected insect cells.
  • The recombinant proteins were then used as antigens in the indirect ELISA. Indirect ELISA is an antibody test that can detect and quantify specific antibodies in a sample.
  • They tested sera from naïve (unvaccinated, unexposed), vaccinated and infected horses. The horses were either from a Hendra-virus free region, or had been pre-tested in validated diagnostic tests.

Results

  • The data revealed the credibility of this approach, as HeV-N-specific antibodies were only detected in sera from infected horses, while HeV-G-specific antibodies were detected in infected and vaccinated horses.
  • The test exhibited a high level of specificity and sensitivity. Specificity refers to the test’s ability to correctly identify those without the disease (true negative rate), while sensitivity relates to the test’s ability to properly identify those with the disease (true positive rate).
  • A significant correlation was noted between data from German and Australian HeV-negative horses, indicating that the developed test could be employed successfully for horse serum samples from diverse geographical regions.

Conclusion

  • The indirect ELISA-based “Differentiating Infected from Vaccinated Animals” (DIVA) approach using recombinant HeV-G and HeV-N proteins can reliably differentiate between horses that are vaccinated against Hendra virus and those who are infected with it.

Cite This Article

APA
Balkema-Buschmann A, Fischer K, McNabb L, Diederich S, Singanallur NB, Ziegler U, Keil GM, Kirkland PD, Penning M, Sadeghi B, Marsh G, Barr J, Colling A. (2022). Serological Hendra Virus Diagnostics Using an Indirect ELISA-Based DIVA Approach with Recombinant Hendra G and N Proteins. Microorganisms, 10(6), 1095. https://doi.org/10.3390/microorganisms10061095

Publication

Microorganisms
ISSN: 2076-2607
NlmUniqueID: 101625893
Country: Switzerland
Language: English
Volume: 10
Issue: 6
PII: 1095

Researcher Affiliations

Balkema-Buschmann, Anne
  • Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
Fischer, Kerstin
  • Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
McNabb, Leanne
  • Australian Centre for Disease Preparedness, CSIRO, Geelong, VIC 3220, Australia.
Diederich, Sandra
  • Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
Singanallur, Nagendrakumar Balasubramanian
  • Australian Centre for Disease Preparedness, CSIRO, Geelong, VIC 3220, Australia.
  • OIE Collaborating Centre for Diagnostic Test Validation Science, Commonwealth Scientific and Industrial Research Organisation, Geelong, VIC 3220, Australia.
Ziegler, Ute
  • Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
Keil, Günther M
  • Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
Kirkland, Peter D
  • Elizabeth Macarthur Agriculture Institute, Menangle, NSW 2567, Australia.
Penning, Maren
  • Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
Sadeghi, Balal
  • Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
Marsh, Glenn
  • Australian Centre for Disease Preparedness, CSIRO, Geelong, VIC 3220, Australia.
Barr, Jennifer
  • Australian Centre for Disease Preparedness, CSIRO, Geelong, VIC 3220, Australia.
Colling, Axel
  • Australian Centre for Disease Preparedness, CSIRO, Geelong, VIC 3220, Australia.
  • OIE Collaborating Centre for Diagnostic Test Validation Science, Commonwealth Scientific and Industrial Research Organisation, Geelong, VIC 3220, Australia.

Grant Funding

  • JA00093092 / Organisation For Economic Co-Operation and Development

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 4 times.
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  3. Mbu'u CM, Gontao P, Wade A, Penning M, Sadeghi B, Mbange AE, LeBreton M, Kamdem SLS, Stoek F, Groschup MH, Mbacham WF, Balkema-Buschmann A. Serological and molecular analysis of henipavirus infections in synanthropic fruit bat and rodent populations in the Centre and North regions of Cameroon (2018-2020). BMC Vet Res 2025 Feb 24;21(1):93.
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