Development of an Immunochromatographic Strip for Rapid Detection of Mink Enteritis Virus.

Jianke Wang,Zhenjun Wang,Li Yi,Yuening Cheng,Shipeng Cheng,Shanshan Song,Peng Lin

doi:10.3389/fmicb.2022.839320

Abstract

Although mink enteritis virus (MEV) is an acute, virulent, and highly contagious pathogen in minks, there is currently a lack of a quick diagnostic method. By conjugating colloidal gold nanoparticles with the MEV-specific monoclonal antibody, monoclonal antibody (MAb) 14, we developed a single-step competitive immunochromatographic strip (ICS) assay for simple determination of MEV. The optimal concentrations of the colloidal gold-coupled MAb 14 (coating antibody), the capture protein (MEV VP2 protein), and the goat anti-mouse antibody were 1.0, 0.8, and 1.0 mg/ml, respectively. The limit of detection was approximately 512 hemagglutination units/100 μl of MEV B strain. Other common viruses of mink were tested to evaluate the specificity of the ICS, and the results showed no cross-reactivity for other pathogens. In comparison with the Anigen Rapid canine parvovirus (CPV) Ag Test Kit (BioNote, Korea) in testing 289 samples, the percentage of agreement and relative sensitivity and specificity of the MEV ICS assay were 94.1, 93.2, and 97.1%, respectively. The ICS test was found to be a sufficiently sensitive and specific detection method for the convenient and rapid detection of MEV.

Highlights

Mink enteritis virus (MEV) can cause acute, virulent, and highly contagious disease in minks and is characterized by acute hemorrhagic enteritis and leucopenia, especially in younger animals
The mink enteritis virus (MEV) B strain (MEVB), canine distemper virus (CDV)-3, canine parvovirus (CPV)-BJ-21, canine adenovirus (CAV)-2c, and Aleutian mink disease virus (AMDV) strains were obtained from the Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences (Lin et al, 2018)
The results of the immunofluorescence assay (IFA) showed that all five monoclonal antibody (MAb) reacted with the MEV-infected F81 cells (Figure 2A), while no fluorescent signal was visualized in negative control with SP2/0 cells suspension

Summary

Introduction

Mink enteritis virus (MEV) can cause acute, virulent, and highly contagious disease in minks and is characterized by acute hemorrhagic enteritis and leucopenia, especially in younger animals. The virus belongs to the genus Protoparvovirus (subfamily, Parvovirinae; family, Parvoviridae). This genus includes canine parvovirus (CPV), feline panleukopenia virus, and raccoon parvovirus. The capsid proteins of these four viruses share 90% similarity (Parrish and Carmichael, 1983). With an average diameter of 18–24 nm, MEV is a nonenveloped ssDNA virus consisting of two large open reading frames (ORFs), which encode two nonstructural proteins (NS1 and NS2) and two structural proteins (VP1 and VP2) through alternative splicing of the mRNAs. VP2 is the main viral capsid, and it comprises approximately 90% of the

Methods

Results

Conclusion