Abstract
Salmonella enteritidis and Salmonella pullorum belonging to Group O9 Salmonella are major causative agents of infectious diseases in chicken. O9 antigen as a part of lipopolysaccharide (LPS) is a predominant detected target for Salmonella infection. To identify the infection, an anti-O9 monoclonal antibody (McAb)-based direct competitive enzyme-linked assay (O9 Dc-ELISA) was developed after constraints were optimized; the establishment and application of O9 Dc-ELISA, compared to two commercial kits and plate agglutination test (PAT), showed that O9 Dc-ELISA could screen out more positive samples than the PAT method could and produce the same agreement rates with commercial kits in terms of sensitivity in addition to strong specificity to clinical serum samples.
Highlights
Salmonella, an important zoonotic pathogen, is one of the major causative agents of food-borne infectious diseases worldwide [1]
A series of dilutions of LPS, horseradish peroxidase (HRP)-labeled O9 monoclonal antibody (McAb), and positive sera were prepared for chessboard titration and optimization (Figure 2)
A standard operating procedure was formulated, after 96-well plates (Biofil company, Canada, FEP101896) were coated with ∼100 μL purified LPS (320 ng/ml) in carbonate bicarbonate buffer (CBS, pH 9.4) at 4◦C for 24 h (Figure 6) and washed with PBST (0.05% Tween 20 in phosphate-buffered saline) two times; 200 μL/well 2% BSA PBS solution was added again for blocking for 3 h at 37◦C, 50 μL 1:2 diluted chicken serum (PBS for blank control) and 50 μL HRP-labeled O9 McAb of 41.6 ng/ml were added at the same time
Summary
Salmonella, an important zoonotic pathogen, is one of the major causative agents of food-borne infectious diseases worldwide [1] Consumption of foods such as egg, chicken, pork, beef, and dairy products contaminated with Salmonella can cause salmonellosis in humans [2,3,4]. This pathogen brings huge economic loss in the animal industry and impacts human health, even death [5,6,7,8]. It is necessary to carry out a seroepidemiological survey on Salmonella for healthy breeding and food safety
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