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Clinical and Vaccine Immunology, September 2009, p. 1360-1365, Vol. 16, No. 9
1071-412X/09/$08.00+0     doi:10.1128/CVI.00148-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Development and Evaluation of an Enzyme-Linked Immunosorbent Assay Based on Recombinant VP2 Capsids for the Detection of Antibodies to Aleutian Mink Disease Virus{triangledown}

Anna Knuuttila,1,2* Pirjo Aronen,3 Auli Saarinen,2,{dagger} and Olli Vapalahti1,2

Division of Microbiology and Epidemiology, Faculty of Veterinary Medicine, P.O. Box 66, 00014 University of Helsinki, Finland,1 Department of Virology, Haartman Institute, P.O. Box 21, 00014 University of Helsinki, Finland,2 Finnish Fur Breeders' Association (STKL), P.O. Box 92, 65101 Vaasa, Finland3

Received 26 March 2009/ Returned for modification 11 May 2009/ Accepted 22 July 2009

Aleutian disease (AD), a common infectious disease in farmed minks worldwide, is caused by Aleutian mink disease virus (AMDV). Serodiagnosis of AD in minks has been based on detection of AMDV antibodies by counterimmunoelectrophoresis (CIE) since the 1980s. The aim of this study was to develop and evaluate an enzyme-linked immunosorbent assay (ELISA) based on recombinant virus-like particles (VLPs) for identifying AMDV antibodies from mink sera. AMDV capsid protein (VP2) of a Finnish wild-type strain was expressed by the baculovirus system in Spodoptera frugiperda 9 insect cells and was shown to self-assemble to VLPs (with an ultrastructure similar to that of the actual virion). A direct immunoglobulin G ELISA was established using purified recombinant AMDV VP2 VLPs as an antigen. Sera from farmed minks were collected to evaluate the AMDV VP2 ELISA (n = 316) and CIE (n = 209) based on AMDV VP2 recombinant antigen in parallel with CIE performed using a commercially available traditional antigen. CIE performed with the recombinant antigen had a sensitivity and specificity of 100% and ELISA a sensitivity of 99% and a specificity of 97%, with reference to CIE performed with the commercial antigen. The results show that the recombinant AMDV VP2 VLPs are antigenic and that AMDV VP2 ELISA is sensitive and specific and encourage further development of the method for high-throughput diagnostics, involving hundreds of thousands of samples in Finland annually.

* Corresponding author. Mailing address: Division of Microbiology and Epidemiology, Faculty of Veterinary Medicine, P.O. Box 66, 00014 University of Helsinki, Finland. Phone: 358 9 191 57049. Fax: 358 9 191 57033. E-mail: anna.knuuttila{at}helsinki.fi

{triangledown} Published ahead of print on 29 July 2009.

{dagger} Present address: Department of Medical Genetics, Haartman Institute, P.O. Box 63, 00014 University of Helsinki, Finland.

Clinical and Vaccine Immunology, September 2009, p. 1360-1365, Vol. 16, No. 9
1071-412X/09/$08.00+0     doi:10.1128/CVI.00148-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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