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Development of PCR Primers for the Detection of Salmonella enterica Serovar Choleraesuis Based on the fliC Gene

机译:基于fliC基因检测肠炎沙门氏菌的PCR引物的开发

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Salmonella enterica serovar Choleraesuis may cause swine salmonellosis and human infection. Because the conventional method for detection of this Salmonella serovar may take 3 to 5 days, a PCR method for detection was evaluated. By comparing the sequence of the phase 1 flagellin (fliC) gene of Salmonella Choleraesuis with that of other Salmonella serovars and of other bacteria species available in GenBank, two PCR primers (flinC-F and flinC-R) were designed. Using these primers, all 97 Salmonella Choleraesuis strains assayed generated the expected PCR product, with a molecular mass of 963 bp. Except for S. enterica Paratyphi C, Salmonella isolates other than Salmonella Choleraesuis and non-Salmonella isolates, including strains of Enterobacteriaceae, all generated negative PCR results. Salmonella Paratyphi C could be differentiated from Salmonella Choleraesuis through the use of primers designed from the viaB gene. When Salmonella Choleraesuis isolates from swine stool, pork, liver, feed, and human whole blood samples were assayed with a preenrichment step, as low as 1 CFU/g or ml of the original sample could be detected.
机译:肠炎沙门氏菌血清霍乱可能引起猪沙门氏菌病和人类感染。由于检测该沙门氏菌血清的常规方法可能需要3至5天,因此对PCR检测方法进行了评估。通过比较霍乱沙门氏菌的1期鞭毛蛋白(fliC)基因与其他沙门氏菌血清型和GenBank中其他细菌种类的序列,设计了两种PCR引物(flinC-F和flinC-R)。使用这些引物,所有97种沙门氏菌霍乱沙门氏菌菌株均产生了预期的PCR产物,分子量为963 bp。除副伤寒沙门氏菌外,除霍乱沙门氏菌和非沙门氏菌以外的沙门氏菌分离株,包括肠杆菌科菌株,均产生阴性PCR结果。通过使用viaB基因设计的引物,可以将副伤寒沙门氏菌与霍乱沙门氏菌区分开。当从猪粪,猪肉,肝脏,饲料和人全血样品中分离出霍乱沙门氏菌时,通过预富集步骤进行分析,可以检测到低至1 CFU / g或ml的原始样品。

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