A network approach for provisional assay recognition of a Hendra virus antibody ELISA: test validation with low sample numbers from infected horses

Axel Colling; Ross Lunt; Jemma Bergfeld; Leanne McNabb; Kim Halpin; Susan Juzva; Kim Newberry; Chris Morrissy; Cameron Loomes; Simone Warner; Ibrahim Diallo; Peter Kirkland; Christopher C Broder; Gemma Carlile; Mai Hlaing Loh; Caryll Waugh; Lynda Wright; James Watson; Debbie Eagles; Kurt Zuelke; Sam McCullough; Peter Daniels

doi:10.1177/1040638718760102

A network approach for provisional assay recognition of a Hendra virus antibody ELISA: test validation with low sample numbers from infected horses

J Vet Diagn Invest. 2018 May;30(3):362-369. doi: 10.1177/1040638718760102. Epub 2018 Feb 21.

Authors

Axel Colling^{1

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6}, Ross Lunt^{1

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6}, Jemma Bergfeld^{1

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3

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6}, Leanne McNabb^{1

2

3

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6}, Kim Halpin^{1

2

3

4

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6}, Susan Juzva^{1

2

3

4

5

6}, Kim Newberry^{1

2

3

4

5

6}, Chris Morrissy^{1

2

3

4

5

6}, Cameron Loomes^{1

2

3

4

5

6}, Simone Warner^{1

2

3

4

5

6}, Ibrahim Diallo^{1

2

3

4

5

6}, Peter Kirkland^{1

2

3

4

5

6}, Christopher C Broder^{1

2

3

4

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6}, Gemma Carlile^{1

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3

4

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6}, Mai Hlaing Loh^{1

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3

4

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6}, Caryll Waugh^{1

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3

4

5

6}, Lynda Wright^{1

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3

4

5

6}, James Watson^{1

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4

5

6}, Debbie Eagles^{1

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3

4

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6}, Kurt Zuelke^{1

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3

4

5

6}, Sam McCullough^{1

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3

4

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6}, Peter Daniels^{1

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6}

Affiliations

¹ CSIRO, Australian Animal Health Laboratory, Newcomb, Victoria, Australia (Colling, Lunt, Bergfeld, McNabb, Halpin, Juzva, Newberry, Morrissy, Carlile, Loh, Waugh, Wright, Watson, Eagles, Zuelke, McCullough, Daniels).
² DAFWA Diagnostic Laboratory Services (DDLS), Animal Pathology, Department of Agriculture and Food, Western Australia, Australia (Loomes).
³ Agribio-Centre for AgriBioSciences, Agriculture Victoria, La Trobe University Campus, Bundoora, Australia (Warner).
⁴ Serology & Virology, Biosecurity Sciences Laboratory, Health and Food Sciences Precinct, Queensland, Australia (Diallo).
⁵ Virology Laboratory, Elizabeth Macarthur Agriculture Institute, Menangle, New South Wales, Australia (Kirkland).
⁶ Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD (Broder).

Abstract

Obtaining statistically sound numbers of sera from Hendra virus (HeV)-infected horses is problematic because affected individuals usually die or are euthanized before developing a serum antibody response. As a consequence, test validation becomes a challenge. Our approach is an extension of OIE principles for provisional recognition and included 7 validation panels tested across multiple laboratories that provided estimates for test performance characteristics. At a 0.4 S/P cutoff, 16 of 19 sera from HeV-infected horses gave positive results in the HeV soluble G, indirect ELISA (HeVsG iELISA; DSe 84.2% [95% CI: 60.4-96.6%]); 463 of 477 non-infected horse sera tested negative (DSp 97.1% [95% CI: 95.1-98.4%]). The HeVsG iELISA eliminated almost all false-positive results from the previously used HeV iELISA, with marginally decreased relative sensitivity. Assay robustness was evaluated in inter-laboratory and proficiency testing panels. The HeVsG iELISA is considered to be fit for purpose for serosurveillance and international movement of horses when virus neutralization is used for follow-up testing of positive or inconclusive serum samples.

Keywords: Hendra virus serology; horses; network; sensitivity; specificity; validation.

Publication types

Evaluation Study
Multicenter Study

MeSH terms

Animals
Antibodies, Viral / blood*
Enzyme-Linked Immunosorbent Assay / veterinary*
Hendra Virus / immunology*
Horse Diseases / virology*
Horses
Sensitivity and Specificity

Substances

Antibodies, Viral