Abstract
Riemerella anatipestifer is one of the most important bacterial pathogen of ducks and causes a contagious septicemia. R. anatipestifer infection causes serositis syndromes similar to other bacterial infections in ducks, including infection by Escherichia coli, Salmonella enterica and Pasteurella multocida. Clinically differentiating R. anatipestifer infections from other bacterial pathogen infections is usually difficult. In this study, MAb 1G2F10, a monoclonal antibody against R. anatipestifer GroEL, was used to develop a colloidal gold immunochromatographic strip. Colloidal gold particles were prepared by chemical synthesis to an average diameter of 20±5.26 nm by transmission electron microscope imaging. MAb 1G2F10 was conjugated to colloidal gold particles and the formation of antibody-colloidal gold conjugates was monitored by UV/Vis spectroscopy. Immunochromatographic strips were assembled in regular sequence through different accessories sticked on PVC plate. Strips specifically detected R. anatipestifer within 10 min, but did not detect E. coli, S. enterica and P. multocida. The detection limit for R. anatipestifer was 1×106 colony forming units, which was 500 times higher than a conventional agglutination test. Accuracy was 100% match to multiplex PCR. Assay stability and reproducibility were excellent after storage at 4°C for 6 months. The immunochromatographic strips prepared in this study offer a specific, sensitive, and rapid detection method for R. anatipestifer, which is of great importance for the prevention and control of R. anatipestifer infections.
Highlights
Riemerella anatipestifer is a Gram-negative, non-motile, rod-shaped bacterium that is one of the most important bacterial pathogen of ducks
R. anatipestifer infection causes a contagious septicemia characterized by fibrinous pericarditis, airsacculitis, perihepatitis, and caseous salpingitis
Recent molecular biological methods include polymerase chain reaction (PCR) [7], multiplex PCR (m-PCR) [8] assay, and loop-mediated isothermal amplification (LAMP) [9] assays; these procedures are very cumbersome since they require skilled technicians, special equipments and reagents, and several hours to perform
Summary
Riemerella anatipestifer is a Gram-negative, non-motile, rod-shaped bacterium that is one of the most important bacterial pathogen of ducks. R. anatipestifer infection causes a contagious septicemia characterized by fibrinous pericarditis, airsacculitis, perihepatitis, and caseous salpingitis. At least 21 serotypes of R. anatipestifer have been identified [1,2,3], with serotypes 1, 2, and 10 are the most prevalent in China [4]. Colloidal Gold Chromatographic Strips for R. anatipestifer Detection
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