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基于 fliC 基因的猪霍乱沙门氏菌血清型 PCR 检测引物的研制。

Development of PCR primers for the detection of Salmonella enterica serovar Choleraesuis based on the fliC gene.

机构信息

Department of Food Science, National Chung-Hsing University, Taichung, Taiwan, Republic of China.

出版信息

J Food Prot. 2005 Aug;68(8):1575-80. doi: 10.4315/0362-028x-68.8.1575.

Abstract

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.

摘要

猪霍乱沙门氏菌可引起猪沙门氏菌病和人类感染。由于传统的沙门氏菌血清型检测方法可能需要 3 到 5 天,因此评估了一种用于检测的 PCR 方法。通过比较猪霍乱沙门氏菌相 1 鞭毛(fliC)基因的序列与 GenBank 中其他沙门氏菌血清型和其他细菌种属的序列,设计了两个 PCR 引物(flinC-F 和 flinC-R)。使用这些引物,检测的 97 株猪霍乱沙门氏菌均产生了预期的 PCR 产物,分子量为 963 bp。除了副伤寒 C 型沙门氏菌外,除了猪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。通过使用来自 viaB 基因的引物可以将副伤寒 C 型沙门氏菌与猪霍乱沙门氏菌区分开来。当对来自猪粪便、猪肉、肝脏、饲料和人全血样本的猪霍乱沙门氏菌分离株进行预富集步骤检测时,可检测到原始样本中低至 1 CFU/g 或 ml 的菌。

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