A fast and robust method for whole genome sequencing of the Aleutian Mink Disease Virus (AMDV) genome

https://doi.org/10.1016/j.jviromet.2016.03.010Get rights and content
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Highlights

  • Method for robust, reliable, and fast sequencing of the AMDV genome.

  • Three step protocol; purification, specific amplification, and Ion-Torrent sequencing.

  • Identification of molecular differences between low and high virulent AMDV strains.

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.

Keywords

AMDV
PCR
NGS
Whole genome sequencing

Cited by (0)

1

Current address: Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Hvidovre Hospital and Department of International Health, Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.

2

Current address: Novo Nordisk A/S, Smørmosevej 17-19, DK-2880 Bagsværd, Denmark.