Sharma V K, Carlson S A
Pre-Harvest Food Safety and Enteric Diseases Research Unit, National Animal Disease Center, Agricultural Research Service, USDA, Ames, Iowa 50010, USA.
Appl Environ Microbiol. 2000 Dec;66(12):5472-6. doi: 10.1128/AEM.66.12.5472-5476.2000.
A multiplex fluorogenic PCR assay for simultaneous detection of pathogenic Salmonella strains and Escherichia coli O157:H7 was developed and evaluated for use in detecting very low levels of these pathogens in meat and feces. Two sets of primers were used to amplify a junctional segment of virulence genes sipB and sipC of Salmonella and an intragenic segment of gene eae of E. coli O157:H7. Fluorogenic reporter probes were included in the PCR assay for automated and specific detection of amplified products. The assay could detect <10 CFU of Salmonella enterica serovar Typhimurium or E. coli O157:H7 per g of meat or feces artificially inoculated with these pathogens and cultured for 6 to 18 h in a single enrichment broth. Detection of amplification products could be completed in </=4 h after enrichment.
开发了一种多重荧光定量PCR检测方法,用于同时检测致病性沙门氏菌菌株和大肠杆菌O157:H7,并评估其在检测肉类和粪便中极低水平的这些病原体方面的应用。使用两组引物扩增沙门氏菌毒力基因sipB和sipC的连接片段以及大肠杆菌O157:H7基因eae的基因内片段。荧光报告探针被纳入PCR检测中,用于自动和特异性检测扩增产物。该检测方法能够检测每克人工接种这些病原体并在单一富集肉汤中培养6至18小时的肉类或粪便中<10 CFU的肠炎沙门氏菌鼠伤寒血清型或大肠杆菌O157:H7。富集后≤4小时即可完成扩增产物的检测。