Abstract
Salmonella has been known as an important zoonotic pathogen that can cause a variety of diseases in both animals and humans. Poultry are the main reservoir for the Salmonella serovars Salmonella Pullorum (S. Pullorum), Salmonella Gallinarum (S. Gallinarum), Salmonella Enteritidis (S. Enteritidis), and Salmonella Typhimurium (S. Typhimurium). The conventional serotyping methods for differentiating Salmonella serovars are complicated, time-consuming, laborious, and expensive; therefore, rapid and accurate molecular diagnostic methods are needed for effective detection and prevention of contamination. This study developed and evaluated a TaqMan multiplex real-time PCR assay for simultaneous detection and differentiation of the S. Pullorum, S. Gallinarum, S. Enteritidis, and S. Typhimurium. In results, the optimized multiplex real-time PCR assay was highly specific and reliable for all four target genes. The analytical sensitivity corresponded to three colony-forming units (CFUs) for these four Salmonella serovars, respectively. The detection limit for the multiplex real-time PCR assay in artificially contaminated samples was 500 CFU/g without enrichment, while 10 CFU/g after pre-enrichment. Moreover, the multiplex real-time PCR was applied to the poultry clinical samples, which achieved comparable results to the traditional bacteriological examination. Taken together, these results indicated that the optimized TaqMan multiplex real-time PCR assay will be a promising tool for clinical diagnostics and epidemiologic study of Salmonella in chicken farm and poultry products.
Highlights
Salmonella is an important zoonotic pathogen that can cause a variety of diseases in both animals and humans (Majowicz et al, 2010)
The Salmonella, Escherichia coli, and Pseudomonas aeruginosa strains were cultured at 37°C in Luria-Bertani (LB)
The genomic DNA extracted from bacterial culture dilutions was used as templates for multiplex real-time PCR
Summary
Salmonella is an important zoonotic pathogen that can cause a variety of diseases in both animals and humans (Majowicz et al, 2010). Detection and differentiation of these Salmonella serovars in poultry farms are required to prevent, control, and eliminate the spread of Salmonella. Gallinarum but not other Salmonella serovars (Kang et al, 2011) Based on these specific gene, PCR assays were applied for detecting different Salmonella serovars (Shah et al, 2005; Kim et al, 2006; Hong et al, 2008; Kang et al, 2011; Xu et al, 2018). This study attempted to develop a rapid multiplex RT-PCR assay for the simultaneous detection and differentiation of the prevalent S
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
