Development of a Peptide ELISA for the Diagnosis of Aleutian Mink Disease

Abstract

Aleutian disease (AD) is a common immunosuppressive disease in mink farms world-wide. Since the 1980s, counterimmunoelectrophoresis (CIEP) has been the main detection method for infection with the Aleutian Mink Disease Virus (AMDV). In this study, six peptides derived from the AMDV structural protein VP2 were designed, synthesized, and used as ELISA antigens to detect anti-AMDV antibodies in the sera of infected minks. Serum samples were collected from 764 minks in farms from five different provinces, and analyzed by both CIEP (a gold standard) and peptide ELISA. A peptide designated P1 (415 aa–433 aa) exhibited good antigenicity. A novel ELISA was developed using ovalbumin-linked peptide P1 to detect anti-AMDV antibodies in mink sera. The sensitivity and specificity of the peptide ELISA was 98.0% and 97.5%, respectively. Moreover, the ELISA also detected 342 early-stage infected samples (negative by CIEP and positive by PCR), of which 43.6% (149/342) were true positives. These results showed that the peptide ELISA had better sensitivity compared with CIEP, and therefore could be preferable over CIEP for detecting anti-AMDV antibodies in serological screening.

Evaluation of an in-house indirect enzyme-linked immunosorbent assay of feline panleukopenia VP2 subunit antigen in comparison to hemagglutination inhibition assay to monitor tiger antibody levels by Bayesian approach

“…Previous studies have identified the capsid protein VP2 of FPV as a coating antigen for ELISA in the detection of antibodies against feline panleukopenia in cats [12]. In addition, the VP2 subunit protein exhibited good antigenicity in ELISA development for the detection of antibodies against parvovirus in minks causing Aleutian Mink disease with high sensitivity and specificity [20]. On that basis, the amino acids 545-585 of VP2, which covered the epitope region and were specific to the FPV, were selected and employed as an antigen in an indirect ELISA test in this study.…”

AMDV Vaccine: Challenges and Perspectives

“…In recent times, this approach has been shown to be successful in reducing the prevalence of infected mink in Nova Scotia Canada, but has failed in eradicating AMDV from most of the infected farms [ 57 ]. Despite its advantages, CIEP is not a very sensitive diagnostic technique, can yield false negative results and is very time consuming [ 58 , 59 ]. In contrast, the enzyme-linked immunosorbent assay (ELISA) is another approach that is frequently used, which is based on detecting anti-AMDV antibodies in blood with the advantage of being automated and more sensitive [ 60 ].…”

A synthetic peptide analog of in silico‐predicted immunogenic epitope unique to dengue virus serotype 2 NS1 antigen specifically binds immunoglobulin G antibodies raised in rabbits

“…One way to solve cross‐reactivity problems in diagnostics is to identify immunogenic epitopes exclusive to a specific pathogen . In previous studies, peptides corresponding to identified epitopes of antigenic markers of infections have been used in diagnostics …”