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Enterotoxigenic profiles and polymerase chain reaction detection of Bacillus cereus group cells and B. cereus strains from foods and food-borne outbreaks

机译:食品和食源性暴发中蜡状芽孢杆菌群细胞和蜡状芽孢杆菌菌株的肠毒理学概况和聚合酶链反应检测

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摘要

Bacillus cereus is one of the important food pathogens. Since B. cereus group cells, such as B. cereus, B. thuringiensis, B. anthracis and B. mycoides, share many phenotypical properties and a high level of chromosomal sequence similarity, it is interesting to investigate the virulence profiles for B. cereus group cells, including B. cereus strains isolated from foods and samples associated with food-poisoning outbreaks. For this investigation, the presence of enterotoxin genes, such as those of haemolysin BL, B. cereus enterotoxin T and enterotoxin FM, were assayed by polymerase chain reaction (PCR) methods. Meanwhile, their enterotoxin activities were assayed using the BCET-RPLA kit, haemolytic patterns on sheep blood agar and their cytotoxicity to Chinese hamster ovary (CHO) cells. Results showed that there were 12 enterotoxigenic profiles for the 98 B. cereus group strains collected. In addition, if any of the three types of enterotoxins was present in the B. cereus group cells, these cells were shown to be cvtotoxic to the CHO cells. Similar enterotoxigenic profiles could be found among strains of B. cereus, B. mycoides and B. thuringiensis. Thus, all B. cereus group strains may be potentially toxigenic and the detection of these cells in foods is important. We thus designed PCR primers, termed Ph1/Ph2, from the sphingomyelinase gene of B. cereus cells. These primers were specific for all B. cereus group strains and could be used for the detection of B. cereus cells contaminated in food samples.
机译:蜡状芽孢杆菌是重要的食物病原体之一。由于蜡状芽孢杆菌组细胞,如蜡状芽孢杆菌,苏云金芽孢杆菌,炭疽芽孢杆菌和霉菌芽孢杆菌,具有许多表型特性和高度的染色体序列相似性,因此研究蜡状芽孢杆菌的毒力谱非常有趣。组细胞,包括从食物和与食物中毒暴发有关的样品中分离的蜡状芽孢杆菌菌株。为了进行这项研究,通过聚合酶链反应(PCR)方法测定了肠毒素基因的存在,例如溶血素BL,蜡状芽孢杆菌肠毒素T和肠毒素FM。同时,使用BCET-RPLA试剂盒,绵羊血琼脂上的溶血模式及其对中国仓鼠卵巢(CHO)细胞的细胞毒性来测定其肠毒素活性。结果显示,收集到的98种蜡状芽孢杆菌组菌株有12种产肠毒素的特征。另外,如果蜡状芽孢杆菌组细胞中存在三种肠毒素中的任何一种,则表明这些细胞对CHO细胞具有细胞毒性。在蜡状芽孢杆菌,霉菌芽孢杆菌和苏云金芽孢杆菌的菌株之间可以发现相似的产毒曲线。因此,蜡状芽孢杆菌的所有菌株都可能具有潜在的毒性,食品中这些细胞的检测很重要。因此,我们从蜡状芽孢杆菌细胞的鞘磷脂酶基因设计了PCR引物,称为Ph1 / Ph2。这些引物对所有蜡状芽孢杆菌属菌株均具有特异性,可用于检测食品样品中污染的蜡状芽孢杆菌细胞。

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