Abstract
The objective of this study was to assess the sensitivity of the Omni Klentaq-LA DNA polymerase for detecting Aleutian mink disease virus (AMDV) in mink blood and tissues by PCR without DNA extraction. The presence of AMDV DNA was directly tested by Klentaq in the plasma, serum, whole blood, and spleen homogenates of 188 mink 4 and 16 months after inoculation with the virus. Samples from bone marrow, small intestine, liver, lungs, kidneys, and lymph nodes of 20 of the same mink were also tested by Klentaq. DNA was extracted from paired samples of plasma and the aforesaid tissues by a commercial nucleic acid extraction kit (Dynabeads Silane) and tested by PCR. Compared with the extracted DNA, Klentaq detected a significantly greater number of samples in the whole blood, serum, plasma, spleen, and small intestine. It was concluded that Klentaq is a preferred system for directly detecting AMDV DNA in mink blood and tissues. The lower success rate of extracted DNA compared with Klentaq could be the result of DNA losses during the extraction process. This is an important factor in chronically infected mink, which have a low AMDV copy number in the bloodstream. Direct AMDV detection also reduces the cost of PCR amplification and lowers the risk of sample contamination.
Highlights
Aleutian mink disease virus (AMDV, Carnivore amdoparvovirus 1), a species of the genus Amdoparvovirus, family Parvoviridae (Cotmore et al 2014), exists in almost all mink-producing countries
The results suggested that the type of organ influenced the chance of PCR success by direct amplification by the Klentaq and by Dynabeads Silane (DB), and that the Klentaq significantly increased the chance of PCR success for spleen, small intestine, and lung samples
DNA extracted from leukocytes provided a larger number of PCR-positive results than that extracted from plasma for the detection of Epstein– Barr virus (Ito et al 2016) and cytomegalovirus (Deback et al 2007; Chen et al 2014)
Summary
Aleutian mink disease virus (AMDV, Carnivore amdoparvovirus 1), a species of the genus Amdoparvovirus, family Parvoviridae (Cotmore et al 2014), exists in almost all mink-producing countries. The virus causes Aleutian disease, an immune-complex syndrome (reviewed in Bloom et al 1994), leading to reduced reproductive performance (Broll and Alexandersen 1996; Reichert and Kostro 2014) and increased adult and kit mortality (Bloom et al 1994; Dyer et al 2000). PCR, on the contrary, can detect infection earlier than CIEP (Jackson et al 1992; Oie et al 1996; Farid et al 2015), which is important when an AMDV-free ranch becomes infected and animals that carry the virus ought to be identified and eliminated as fast as possible. PCR fails to detect the virus in some chronically infected mink when virus replication becomes restricted (Jackson et al 1996; Jensen et al 2014)
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