Development of a colloidal gold immunochromatographic strip for rapid detection of Riemerella anatipestifer in ducks

Abstract

Riemerella anatipestifer is one of the major bacterial pathogens of ducks and causes significant economic losses in poultry agriculture. Usually, methods for detecting R. anatipestifer infection need specialized equipment and highly skilled personnel. In this study, a novel colloidal gold immunochromatographic strip was developed for rapid detection of R. anatipestifer in ducks. The monoclonal antibodies 2D5 and 2A6 against R. anatipestifer were used as colloidal gold-labeled protein and capture protein, respectively, to recognize the bacteria in tryptic soy broth medium culture and in hearts of infected ducks. The goat anti-mouse IgG antibody was labeled on nitrocellulose membrane as a control for C line. The labeling pH was optimized as 10.0, and the concentration of 2D5 labeled to colloidal gold particles was optimized as 18 μg/mL. The strip specifically detected serotypes 1, 2, and 10 R. anatipestifer strains and showed no cross-reaction with Escherichia coli, Salmonella enterica, and Pasteurella multocida strains. The sensitivity of the strip for detecting R. anatipestifer was 1.0 × 106 colony forming unit. The strips remained stable for up to 8 mo at 4°C, and the detection can be completed within 15 min. The strip can detect R. anatipestifer in hearts of the ducks experimentally infected with R. anatipestifer but not infected with E. coli, which were also confirmed with bacterial isolation followed by multiplex polymerase chain reaction. These results suggested that the strips are reliable methods for identification of R. anatipestifer in laboratories and in duck farms.

Keywords

• Riemerella anatipestifer
• colloidal gold
• monoclonal antibody 2D5
• monoclonal antibody 2A6
• immunochromatographic strip