Abstract
BackgroundSalmonella enterica serovar Pullorum is a host-restricted serotype causing infection in poultry. The pathogen can not only cause acute infection in young chicks with high mortality and morbidity, but also persist in adult chickens without evident clinical symptoms and lead to vertical transmission. To eradicate S. Pullorum in poultry farms, it is necessary to establish an efficient method to monitor the prevalence of the pathogen in adult chickens. The protein IpaJ is a specific immunogen in S. Pullorum and is not detected in closely related serotypes, such as S. Gallinarum and S. Enteritidis.ResultsIn the present study, IpaJ was expressed as a recombinant fusion protein MBP-IpaJ in E. coli. The purified MBP-IpaJ was used as a coating antigen to develop an indirect ELISA assay, which was applied to the detection of S. Pullorum infection in chickens. The indirect ELISA assay demonstrated that antibodies produced against IpaJ were detectable in antisera of chickens infected with S. Pullorum in the second week, stably increased until the tenth week, and persisted at a high level in the following two weeks. Furthermore, the ELISA method detected four positive samples out of 200 clinical antiserum samples collected from a poultry farm, and the positive samples were confirmed to be reacted with S. Pullorum using the standard plate agglutination test.ConclusionsThe established indirect ELISA using the IpaJ protein is a novel method for specific detection of S. Pullorum infection, and contribute to eradication of pullorum disease in the poultry industry.
Highlights
Salmonella enterica serovar Pullorum is a host-restricted serotype causing infection in poultry
The recombinant strain ER2523-pMAL-c5X-ipaJ was subjected to induction with 0.3 mM IPTG, and SDS-PAGE results showed that Maltose binding protein (MBP)-IpaJ was successfully expressed in the host cells (Fig. 1b)
The MBP-IpaJ fusion protein was expressed in the supernatant and successfully purified using affinity chromatography (Fig. 1c)
Summary
Salmonella enterica serovar Pullorum is a host-restricted serotype causing infection in poultry. It is necessary to establish an efficient method to monitor the prevalence of the pathogen in adult chickens. Pullorum), a host-restricted serotype mainly infecting poultry [1]. To reduce the prevalence of the disease in poultry farms, efficient surveillance has become increasingly important and Serology detection has frequently been used to identify infected and carrier animals in both surveillance and epidemiological studies [6]. The gold standard serological Salmonella detection method in chickens is the agglutination test, which played a significant role in (2019) 15:3 eradicating S. ELISA methods were subsequently developed to detect antibodies against LPS or FliC in avian sera [8,9,10,11], but problems of cross-reactivity could not be resolved because of antigen conservation in the different serotypes. Pullorum infection is useful for surveillance in the poultry industry
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