Abstract
BackgroundAleutian 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).MethodsA 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.ResultsThe 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%.ConclusionsThe 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.
Highlights
Aleutian mink disease virus (AMDV) is the cause of a chronic immune complex disease, Aleutian disease (AD), which is common in mink-producing countries
Animals and herds We sampled and screened for anti-AMDV antibodies on two mink farms and used both CIEP with capillary blood samples, and the automated enzyme-linked immunosorbent assay (ELISA) system with recombinant Structural protein 2 (VP2) antigen and filter-paper (FP) strips
Our aim was to automate and validate an ELISA system for detecting anti-AMDV antibodies in the blood of farmed mink
Summary
Aleutian mink disease virus (AMDV) is the cause of a chronic immune complex disease, Aleutian disease (AD), which is common in mink-producing countries. Aleutian disease (AD), a common and economically significant disease in farmed mink (Neovison vison), is caused by Aleutian mink disease virus (AMDV). In 2005, the Finnish Fur Breeders’ Association (FFBA) implemented an eradication programme to depopulate AMDV-infected farms and reduce the overall prevalence of AMDV, and to improve the health status of animals and reduce economic losses to farmers caused by AD. In groups B, C, and D, withinfarm prevalence should not exceed 1, 2, and 50 positive/ 1000 breeding females, respectively. In group E, prevalence exceeds 50 positive/1000 breeding females. Fin Furlab (formerly Fur Animal Feed Laboratory) tests 500,000 to 600,000 samples annually for anti-AMDV antibodies. From 2000 to 2012, the annual mean seroprevalence of all tested animals ranged between 3.3% and 13.6%
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