tailieunhanh - Báo cáo khoa học: "A multiplex real-time PCR for differential detection and quantification of Salmonella spp., Salmonella enterica serovar Typhimurium and Enteritidis in meats"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: A multiplex real-time PCR for differential detection and quantification of Salmonella spp., Salmonella enterica serovar Typhimurium and Enteritidis in meats | J. Vet. Sci. 2009 10 1 43-51 DOI JOURNAL OF Veterinary Science A multiplex real-time PCR for differential detection and quantification of Salmonella spp. Salmonella enterica serovar Typhimurium and Enteritidis in meats 1 1 2 1 1 1 Su Hwa Lee Byeong Yeal Jung Nabin Rayamahji Hee Soo Lee Woo Jin Jeon Kang Seuk Choi Chang Hee Kweon1 Han Sang Yoo2 National Veterinary Research and Quarantine Service Anyang 430-824 Korea 2Department of Infectious Diseases College of Veterinary Medicine KRF Priority Zoonotic Disease Research Institute and BK21 Program for Veterinary Science Seoul National University Seoul 151-742 Korea Salmonella S. Typhimurium and S. Enteritidis are the major causative agents of food-borne illnesses worldwide. Currently a rapid detection system using multiplex real-time polymerase chain reaction PCR has been applied for other food-borne pathogens such as Escherichia coli Staphylococcus aureus and Streptococcus spp. A multiplex real-time PCR was developed for the simultaneous detection of Salmonella spp. especially S. Typhimurium and S. Enteritidis in beef and pork. For the specific and sensitive multiplex real-time PCR three representative primers and probes were designed based on sequence data from Genbank. Among the three DNA extraction methods boiling alkaline lysis and QIAamp DNA Mini Kit the QIAamp DNA Mini Kit was the most sensitive in this study. The optimized multiplex real-time PCR was applied to artificially inoculated beef or pork. The detection sensitivity of the multiplex real-time PCR was increased. The specificity of the multiplex real-time PCR assay using 128 pure-cultured bacteria including 110 Salmonella isolates and 18 nonSalmonella isolates was 100 100 and for Salmonella spp. S. Typhimurium and S. Enteritidis respectively. The sensitivity was 100 100 and for Salmonella spp. S. Typhimurium and S. Enteritidis respectively. The multiplex real-time PCR assay developed in this study could detect up to .