title: A fast and robust method for whole genome sequencing of the Aleutian Mink Disease Virus (AMDV) genome.
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.
authors.affiliation: Kopenhagen Diagnostics, Kopenhagen Fur, Glostrup, Denmark; Department of Systems biology, Technical University of Denmark, Lyngby, Denmark. Electronic address: eha@kopenhagenfur.com.
authors.forename: Emma E
authors.initials: EE
authors.lastname: Hagberg
authors.affiliation: Kopenhagen Diagnostics, Kopenhagen Fur, Glostrup, Denmark.
authors.forename: Anders
authors.initials: A
authors.lastname: Krarup
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark.
authors.forename: Ulrik
authors.initials: U
authors.lastname: Fahnøe
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark.
authors.forename: Lars E
authors.initials: LE
authors.lastname: Larsen
authors.affiliation: Kopenhagen Diagnostics, Kopenhagen Fur, Glostrup, Denmark.
authors.forename: Rebekka
authors.initials: R
authors.lastname: Dam-Tuxen
authors.affiliation: Department of Systems biology, Technical University of Denmark, Lyngby, Denmark.
authors.forename: Anders G
authors.initials: AG
authors.lastname: Pedersen
mesh_terms: D000454:Aleutian Mink Disease Virus / Q000235:genetics* / Q000302:isolation & purification; D000818:Animals; D004279:DNA, Viral / Q000235:genetics; D016679:Genome, Viral; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D010802:Phylogeny; D016133:Polymerase Chain Reaction
chemical_list: D004279:DNA, Viral
keywords: AMDV; NGS; PCR; Whole genome sequencing