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Open Access Research

Validation of an automated ELISA system for detection of antibodies to Aleutian mink disease virus using blood samples collected in filter paper strips

Anna Knuuttila12*, Pirjo Aronen3, Majvor Eerola3, Ian A Gardner4, Anna-Maija K Virtala1 and Olli Vapalahti12

Author Affiliations

1 Department of Veterinary Biosciences, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland

2 Department of Virology, Haartman Institute, Faculty of Medicine, University of Helsinki, Helsinki, Finland

3 Fin Furlab Oy, Finnish Fur Breeders’ Association, Vaasa, Finland

4 Department of Health Management, University of Prince Edward Island, Charlottetown, PE, Canada

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Virology Journal 2014, 11:141  doi:10.1186/1743-422X-11-141

Published: 8 August 2014

Abstract

Background

Aleutian mink disease virus (AMDV) is the cause of a chronic immune complex disease, Aleutian disease (AD), which is common in mink-producing countries. In 2005, implementation of an AMDV eradication programme in Finland created a need for an automated high-throughput assay. The aim of this study was to validate an AMDV-VP2 -recombinant antigen ELISA, which we developed earlier, in an automated assay format for the detection of anti-AMDV antibodies in mink blood and to determine the accuracy of this test compared with the reference standard (counter-current immunoelectrophoresis, CIEP).

Methods

A blood sampling method based on filter paper 12-strips (blood combs) and a device to introduce these strips to an ELISA plate for elution of the samples were developed. Blood and serum samples were collected from 761 mink from two farms with low (2%) and high (81%) seroprevalences of AMDV infection in 2008. ELISA sensitivity and specificity were estimated with a Bayesian 2-test 2-population model that allowed for conditional dependence between CIEP and ELISA. Agreement between the two tests was assessed with kappa statistic and proportion agreement.

Results

The sensitivity and specificity of the automated ELISA system were estimated to be 96.2% and 98.4%, respectively. Agreement between CIEP and ELISA was high, with a kappa value of 0.976 and overall proportion agreement of 98.8%.

Conclusions

The automated ELISA system combined with blood comb sampling is an accurate test format for the detection of anti-AMDV antibodies in mink blood and offers several advantages, including improved blood sampling and data handling, fast sample throughput time, and reductions in costs and labour inputs.

Keywords:
Aleutian mink disease virus; Bayesian analysis; Carnivore amdoparvovirus 1; Counter-current immunoelectrophoresis; Enzyme-linked immunosorbent assay; Sensitivity; Specificity