Abstract
BackgroundDuck enteritis virus (DEV) infection causes substantial economic losses to the worldwide duck-producing areas. The monitoring of DEV-specific antibodies is a key to evaluate the effect of DEV vaccine and develop rational immunization programs. Thus, in this study, an immunochromatographic strip (ICS) test was developed for detecting DEV serum antibodies.ResultsThe ICS test is based on membrane chromatography, and uses both the purified recombinant UL51 protein conjugated with colloidal gold and goat anti-rabbit IgG conjugated with colloidal gold as tracers, the purified recombinant UL51 protein as the capture reagent at the test line, and rabbit IgG as the capture reagent at the control line. The specificity of the ICS was evaluated by sera against DEV, Duck hepatitis virus (DHV), Riemerella anatipestifer (RA), Duck E. coli, Muscovy duck parvovirus (MPV), or Duck Influenza viruses (DIV). Only sera against DEV showed the strong positive results. In order to determine the sensitivity of the ICS, anti-DEV serum diluted serially was tested, and the minimum detection limit of 1:128 was obtained. The ICS components, which are provided in a sealed package, require no refrigeration and are stable for 12 months. To evaluate the effect of the ICS, 110 duck serum samples collected from several non-immune duck flocks were simultaneously tested by the ICS test, enzyme-linked immunosorbent assay (ELISA) and neutralization test (NT). The results showed that the sensitivity of the ICS test was almost consistent with ELISA and much higher than NT, has low cost, and is rapid (15 min) and easy to perform with no requirement of specialized equipment, reagent or technicians.ConclusionsIn this work, we successfully developed a simple and rapid ICS test for detecting DEV serum antibodies for the first time. The ICS test was high specific and sensitive for the rapid detection of anti-DEV antibodies, and has great potential to be used for the serological surveillance of DEV infection in the field.
Highlights
Duck enteritis virus (DEV) infection causes substantial economic losses to the worldwide duckproducing areas
Western blotting using positive rabbit anti-DEV antiserum as the first antibody demonstrated that the recombinant UL51 protein reacted strongly and with the antiserum raised against DEV (Figure 1b), suggesting that the purified recombinant UL51 protein was suitable as the capture reagent of the immunochromatographic strip (ICS)
Specificity, sensitivity and stability of the ICS test All of the 5 healthy ducks serum samples and 25 standard serum samples positive for other non-DEV pathogens were found negative for anti-DEV antibodies with the ICS test (Figure 2)
Summary
Duck enteritis virus (DEV) infection causes substantial economic losses to the worldwide duckproducing areas. In this study, an immunochromatographic strip (ICS) test was developed for detecting DEV serum antibodies. Duck viral enteritis (DVE) is an acute contagious disease of various types of waterfowl (ducks, geese, and swans) caused by duck enteritis virus (DEV), which is a member of the subfamily Alpha-herpesviridae [1]. In duck-producing areas of the world where the disease has been reported, DVE has resulted in significant economic losses in domestic and wild waterfowls due to high mortality, condemnations and decreased egg production [1]. Several studies have indicated that DVE is difficult to monitor and control, because DEV establishes an asymptomatic carrier state in both farmed and wild waterfowl and it is only detectable during the intermittent shedding period of the virus [1,4]
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