Primers specific for the fimbrial major subunit gene stdA can be used to detect Salmonella enterica serovars
Primers specific for the fimbrial major subunit gene stdA can be used to detect Salmonella enterica serovars
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DOI：
10.4315/0362-028x-71.6.1108
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发表时间：
2008-06-01
影响因子：
2
通讯作者：
Yeh, Kuang-Sheng
中科院分区：
文献类型：
--
作者：
Chuang, Yin-Ching;Yang, Chia-Huei;Yeh, Kuang-Sheng
探讨了使用 stdA 基因（编码伤寒沙门氏菌中 std 菌毛基因簇的菌毛主要亚基）内部的两个引物通过 PCR 检测沙门氏菌的可行性。 518 bp stdA 特异性序列在肠沙门氏菌 45 个血清型的 268 个菌株中保守。一种邦戈里沙门氏菌 CCUG 30042 菌株和 34 种非沙门氏菌菌株不具有该序列。敏感性测试显示，stdA 特异性引物组在连续稀释的鼠伤寒沙门氏菌中检测到 3.4 x 10(-1) pg 基因组 DNA 和 3.0 x 10(5) CFU/ml。细胞。还使用被鼠伤寒沙门氏菌污染的猪尸体海绵样品进行了体外特异性测试。在 4.0 x 10(7) CFU/ml 的本地非沙门氏菌群存在的情况下，在 37 ℃ 的缓冲蛋白胨水中预富集 18 小时后，猪尸体渗出物中最初的 4.4 x 10(1) CFU/ml 鼠伤寒沙门氏菌接种物达到了 stdA 引物检测水平，但检测水平下降至 4.4 x 10(0)在 42°C 下在 Rappaport-Vassiliadis R10 肉汤中选择性富集 18 小时后的 CFU/ml。使用 stdA 特异性引物的 PCR 方法是检测肠沙门氏菌的快速而灵敏的工具，而肠沙门氏菌是食源性疾病的重要原因。
The feasibility of using two primers internal to the stdA gene (which encodes the fimbrial major subunit of the std fimbrial gene cluster in Salmonella enterica serovar Typhi) to detect Salmonella by PCR was explored. The 518-bp stdA specific sequence was conserved among 268 strains from 45 serovars of S. enterica. One Salmonella bongori CCUG 30042 strain and 34 non-Salmonella strains did not possess this sequence. A sensitivity test revealed that the stdA-specific primer set detected 3.4 x 10(-1) pg of genomic DNA and 3.0 x 10(5) CFU/ml with serial dilutions of Salmonella Typhimurium. cells. In vitro testing for specificity using pig carcass sponge samples contaminated with Salmonella Typhimurium also was performed. An initial Salmonella Typhimurium inoculum of 4.4 x 10(1) CFU/ml in pig carcass exudates reached the stdA primer detection level after preenrichment in buffered peptone water at 37 degrees C for 18 h in the presence of indigenous non-Salmonella flora at 4.0 x 10(7) CFU/ml, but the detection level decreased to 4.4 x 10(0) CFU/ml after selective enrichment in Rappaport-Vassiliadis R10 broth for 18 h at 42 degrees C. The PCR method with primers specific for stdA is a quick and sensitive tool for detecting S. enterica, which is an important cause of foodborne disease.