ABSTRACT
Aleutian Mink Disease Virus (AMDV) is a frequently encountered pathogen associated with commercial mink breeding. AMDV infection leads to increased mortality and compromised animal health and welfare. Currently little is known about the molecular evolution of the virus, and the few existing studies have focused on limited regions of the viral genome. This paper describes a robust, reliable, and fast protocol for amplification of the full AMDV genome using long-range PCR. The method was used to generate next generation sequencing data for the non-virulent cell-culture adapted AMDV-G strain as well as for the virulent AMDV-Utah strain. Comparisons at nucleotide- and amino acid level showed that, in agreement with existing literature, the highest variability between the two virus strains was found in the left open reading frame, which encodes the non-structural (NS1-3) genes. This paper also reports a number of differences that potentially can be linked to virulence and host range. To the authors' knowledge, this is the first study to apply next generation sequencing on the entire AMDV genome. The results from the study will facilitate the development of new diagnostic tools and can form the basis for more detailed molecular epidemiological analyses of the virus.
Subject(s)
Aleutian Mink Disease Virus/genetics , Genome, Viral , High-Throughput Nucleotide Sequencing/methods , Aleutian Mink Disease Virus/isolation & purification , Animals , DNA, Viral/genetics , Phylogeny , Polymerase Chain ReactionABSTRACT
Aleutian disease (AD) is a severe disease characterized by hypergammaglobulinemia causing multiple symptoms such as acute renal failure, arteritis, reduced reproductive performance and pneumonia in mink. AD is caused by the parvovirus Aleutian mink disease virus (ADV) and diagnosed primarily based on ADV serology sometimes supplemented by organ PCR analysis. In Denmark, approximately 3.5-4 million serum samples are tested every year for the presence of anti ADV antibodies as part of a national eradication program. The present study compares the diagnostic performance of the two most commonly used assays for serological screening for Aleutian disease: counter current immunoelectrophoresis (CIEP) and ELISA. In total, 3810 mink were sampled in doublets and analyzed by CIEP and a newly developed fully automated ELISA. The results show that the two assays have a comparable diagnostic performance with the ELISA having a higher sensitivity but lower specificity than the CIEP assay. The ELISA has been approved by the Danish authorities for diagnosing Aleutian disease in mink.