Development and evaluation of an immunochromatographic strip test based on the recombinant UL51 protein for detecting antibody against duck enteritis virus.
BACKGROUND
Duck 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.


RESULTS
The 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.


CONCLUSIONS
In 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.
Shen
Chanjuan
C
Avian Diseases Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Ya'an, Sichuan, 625014, China.
Cheng
Anchun
A
Wang
Mingshu
M
Sun
Kunfeng
K
Jia
Renyong
R
Sun
Tao
T
Zhang
Na
N
Zhu
Dekang
D
Luo
Qihui
Q
Zhou
Yi
Y
Chen
Xiaoyue
X
D018138:Alphaherpesvirinae / Q000276:immunology*; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D002845:Chromatography / Q000379:methods; D004372:Ducks; D017979:Gold Colloid; D006566:Herpesviridae Infections / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary*; D007118:Immunoassay / Q000379:methods; D011201:Poultry Diseases / Q000276:immunology* / Q000821:virology; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity; D014764:Viral Proteins; D014773:Virology / Q000379:methods*
D000914:Antibodies, Viral; D017979:Gold Colloid; D011994:Recombinant Proteins; D014764:Viral Proteins