Development of a Colloidal Gold Immunochromatographic Assay for Duck Enteritis Virus Detection Using Monoclonal Antibodies.

Fengli Liu,Jun Wang,Yu Huang,Maokai Yan,Qingshui Zhang,Mengxu Sun,Jingliang Su,Yanxin Cao,Guanghua Fu,Rongchang Liu

doi:10.3390/pathogens10030365

Abstract

Duck viral enteritis is a highly contagious and fatal disease of commercial waterfowl flocks. The disease occurs sporadically or epizootically in mainland China due to insufficient vaccinations. Early and rapid diagnosis is important for preventive intervention and the control of epizootic events in clinical settings. In this study, we generated two monoclonal antibodies (MAbs) that specifically recognized the duck enteritis virus (DEV) envelope glycoprotein B and tegument protein UL47, respectively. Using these MAbs, a colloidal gold-based immunochromatographic assay (ICA) was developed for the efficient detection of DEV antigens within 15 min. Our results showed that the detection limit of the developed ICA strip was 2.52 × 103 TCID50/mL for the virus infected cell culture suspension with no cross-reactivity with other pathogenic viruses commonly encountered in commercially raised waterfowl. Using samples from experimentally infected ducks, we demonstrated that the ICA detected the virus in cloacal swab samples on day three post-infection, demonstrating an 80% concordance with the PCR. For tissue homogenates from ducks succumbing to infection, the detection sensitivity was 100%. The efficient and specific detection by this ICA test provides a valuable, convenient, easy to use and rapid diagnostic tool for DVE under both laboratory and field conditions.

Highlights

IntroductionDuck enteritis virus (DEV), named Anatid herpesvirus-1, belongs to the genus
Duck enteritis virus (DEV), named Anatid herpesvirus-1, belongs to the genusMardivirus, subfamily Alphaherpesvirinae, of the family Herpesviridae
Western blotting showed that the two monoclonal antibodies (MAbs) recognized distinct polypeptide bands of the duck enteritis virus (DEV) (Figure 2)

Summary

Introduction

Duck enteritis virus (DEV), named Anatid herpesvirus-1, belongs to the genus. Mardivirus, subfamily Alphaherpesvirinae, of the family Herpesviridae. It is the causative agent for duck viral enteritis (DVE), called duck plague. The DEV virion has four structural components that include the lipid envelope, tegument, capsid and a linear double-stranded DNA genome, which is approximately 158 kb and contains 78 open reading frames predicted to encode potential functional proteins [1,2]. Birds of the family Anatidae such as ducks, geese and swans are highly susceptible to DEV infection, which can be transmitted by direct contact with DEV infected birds or indirectly by contact with a virus-contaminated environment. Early and rapid diagnosis is a key preventive intervention that would reduce the spread of DEV and may control an epidemic

Methods

Results

Conclusion