title: An Efficient Multiplex PCR-Based Assay as a Novel Tool for Accurate Inter-Serovar Discrimination ofSalmonellaEnteritidis,S. Pullorum/Gallinarum andS. Dublin.
abstract: Salmonella entericaserovars Enteritidis, Pullorum/Gallinarum, and Dublin are infectious pathogens causing serious problems for pig, chicken, and cattle production, respectively. Traditional serotyping forSalmonellais costly and labor-intensive. Here, we established a rapid multiplex PCR method to simultaneously identify three prevalentSalmonellaserovars Enteritidis, Pullorum/Gallinarum, and Dublin individually for the first time. The multiplex PCR-based assay focuses on three genestcpS, lygD, andflhB. GenetcpSexists only in the threeSalmonellaserovars, andlygDexists only inS. Enteritidis, while a truncated region offlhBgene is only found inS. Pullorum/Gallinarum. The sensitivity and specificity of the multiplex PCR assay using three pairs of specific primers for these genes were evaluated. The results showed that this multiplex PCR method could accurately identifySalmonellaEnteritidis, Pullorum/Gallinarum, and Dublin from eight non-Salmonellaspecies and 27Salmonellaserovars. The least concentration of genomic DNA that could be detected was 58.5 pg/μL and the least number of cells was 100 CFU. Subsequently, this developed method was used to analyze clinicalSalmonellaisolates from one pig farm, one chicken farm, and one cattle farm. The results showed that blinded PCR testing ofSalmonellaisolates from the three farms were in concordance with the traditional serotyping tests, indicating the newly developed multiplex PCR system could be used as a novel tool to accurately distinguish the three specificSalmonellaserovars individually, which is useful, especially in high-throughput screening.
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Yangzhou UniversityYangzhou, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of EducationYangzhou, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou UniversityYangzhou, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Xiong
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Yangzhou UniversityYangzhou, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of EducationYangzhou, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou UniversityYangzhou, China.
authors.forename: Li
authors.initials: L
authors.lastname: Song
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Yangzhou UniversityYangzhou, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of EducationYangzhou, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou UniversityYangzhou, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Tao
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Yangzhou UniversityYangzhou, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of EducationYangzhou, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou UniversityYangzhou, China.
authors.forename: Huijuan
authors.initials: H
authors.lastname: Zheng
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Yangzhou UniversityYangzhou, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of EducationYangzhou, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou UniversityYangzhou, China.
authors.forename: Zihao
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Yangzhou UniversityYangzhou, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of EducationYangzhou, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou UniversityYangzhou, China.
authors.forename: Shizhong
authors.initials: S
authors.lastname: Geng
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Yangzhou UniversityYangzhou, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of EducationYangzhou, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou UniversityYangzhou, China.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Pan
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Yangzhou UniversityYangzhou, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of EducationYangzhou, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou UniversityYangzhou, China.
authors.forename: Xinan
authors.initials: X
authors.lastname: Jiao
keywords: Salmonella Dublin; Salmonella Enteritidis; Salmonella Pullorum/Gallinarum; accurate discrimination; multiplex PCR