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Implementation and validation of a sensitive PCR detection method in the eradication campaign against Aleutian mink
Trine H Jensen1, Laurids S Christensen, Mariann Chriél
1Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark, Aarhus, Denmark. trje@vet.dtu.dk
Abstract:
Aleutian mink disease virus (AMDV) is a severe progressive disease causing multiple different clinical syndromes in mink. In Denmark, the disease is notifiable and under official control. The control programme, based on serological screening, has confined successfully AMDV to the northern part of Denmark. However, re-infections and new introductions of virus into farms require a confirmatory virological test to verify the positive test results of single animals and ultimately to investigate disease transmission. A one step PCR amplifying a 374-base fragment of the NS1 gene of AMDV was compared to the counter-current immune electrophoresis (CIE) routinely used in the serological screening programme. Mink organs (n=299) obtained from 55 recently infected farms and 8 non-infected farms from 2008 to 2010 were tested by PCR, and the results were found to have a high correlation with the serological status of the mink. The relative diagnostic sensitivity of the PCR was 94.7%, and the relative diagnostic specificity was 97.9% when read in parallel with the CIE. PCR positive samples were sequenced and phylogenetic analysis revealed high similarity within the analysed AMDV strains and to AMDV strains described previously.
Insights
A new PCR test accurately detects Aleutian mink disease virus (AMDV) in mink, offering a vital virological confirmation for disease control and transmission studies.
Area of Science:
- Veterinary Virology
- Molecular Diagnostics
- Animal Disease Surveillance
Background:
- Aleutian mink disease virus (AMDV) causes severe, progressive syndromes in mink.
- AMDV is a notifiable disease under official control in Denmark, primarily managed through serological screening.
- Existing control programs face challenges from re-infections and new virus introductions, necessitating confirmatory virological testing.
Purpose of the Study:
- To evaluate a one-step PCR assay for detecting AMDV in mink organs.
- To compare the diagnostic performance of PCR against the routine counter-current immune electrophoresis (CIE) serological test.
- To investigate AMDV transmission dynamics through virological confirmation.
Main Methods:
- A one-step PCR assay targeting a 374-base fragment of the NS1 gene of AMDV was developed.
- Mink organs (n=299) from infected and non-infected farms (2008-2010) were tested using PCR.
- PCR results were correlated with the serological status of the mink and compared to CIE results.
Main Results:
- The PCR assay demonstrated high diagnostic sensitivity (94.7%) and specificity (97.9%) when compared to CIE.
- PCR results showed a strong correlation with the serological status of the mink.
- Phylogenetic analysis of PCR-positive samples revealed high similarity among analyzed AMDV strains.
Conclusions:
- The developed PCR assay is a sensitive and specific tool for confirming AMDV infections in mink.
- This PCR test can aid in verifying serological results and investigating disease transmission in mink farms.
- The findings support the integration of molecular diagnostics into AMDV control strategies.
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