Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus.

Shujie Wang,Jinying Ge,Tongqing An,Guixin Duan,Xi Li,Yongjun Wen,Kongbin Yang,Xuehui Cai,MingXia Sun

doi:10.3389/fmicb.2019.02882

Abstract

Although canine adenovirus (CAdV) is highly prevalent in dogs, there is currently a lack of a quick diagnostic method. In this study, we developed a rapid immunochromatographic strip (ICS) assay using colloidal gold coupled to CAdV-2-specific monoclonal antibodies (mAbs). BALB/c mice were immunized with a purified CAdV-2 suspension, and four mAbs (belonging to two different epitopes) were generated and designated as 2C1, 7D7, 10D1, and 4G1. Western blot and protein spectral analysis indicated that the hexon protein of CAdV-2 recognized all four mAbs. The colloidal gold-coupled 7D7 and 2C1 mAbs were chosen for inclusion in the rapid ICS assay. The optimal concentrations of the coating antibody (2C1), the capture antibody (7D7), and the goat anti-mouse antibody were 1.0 mg/ml, 10 μg/ml, and 2.0 mg/ml, respectively. The limit of detection was approximately 2.0 × 102 tissue culture infective dose (TCID50)/ml. Other common canine viruses were tested to evaluate the specificity of the ICS, and positive results were observed for only CAdV-1 and CAdV-2. The ICS test was conducted on 360 samples to detect CAdV, and the results were compared with those of polymerase chain reaction (PCR) tests. The ICS test was found to be a sufficiently sensitive and specific detection method for the convenient and rapid detection of CAdV.

Highlights

Canine adenovirus (CAdV; family Adenoviridae; genus Mastadenovirus) is a non-enveloped dsDNA virus that infects numerous mammalian carnivores (Morrison et al, 1997; Balboni et al, 2013)
Serological and pathogen surveys conducted in Thailand, Scandinavia, South Africa, Italy, and Turkey demonstrate the high prevalence of canine adenovirus (CAdV)-2 infections in wild dogs and red foxes (Balboni et al, 2013; Bulut et al, 2013)
immunofluorescence assay (IFA) was performed on CAdV type 2 (CAdV-2)-infected MadinDarby canine kidney (MDCK) cells to assess whether the monoclonal antibodies (mAbs) recognized the wild type virus

Summary

Introduction

Canine adenovirus (CAdV; family Adenoviridae; genus Mastadenovirus) is a non-enveloped dsDNA virus that infects numerous mammalian carnivores (Morrison et al, 1997; Balboni et al, 2013). CAdV-1 primarily causes subclinical infection, but can be epizootic in wild carnivores belonging to the Canidae, Mustelidae and Ursidae families (Hechinger et al, 2017; Dowgier et al, 2018; Balboni et al, 2019). Serological and pathogen surveys conducted in Thailand, Scandinavia, South Africa, Italy, and Turkey demonstrate the high prevalence of CAdV-2 infections in wild dogs and red foxes (Balboni et al, 2013; Bulut et al, 2013)

Methods

Results

Conclusion