DETECTION OF HBLD TOXIN GENE BY BACILLUS CEREUS ISOLATED FROM MEAT CURRY FOOD SAMPLES IN MALAYSIAN RESTAURANTS

MARWAN Jawad MSARAH, Ahmed Alsier, A. Sahilah
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引用次数: 3

Abstract

Bacillus cereus is a ubiquitous foodborne pathogen, can cause food poisoning, leading to infections, have two major types of food poisoning emetic and diarrheal. Foods rich in protein such as meat are associated with foodborne outbreaks of diarrhea caused by B. cereus. The aim of this study is to isolate and identify B. cereus from ready to eat (RTE) meat curry from restaurants in Malaysia and to detect hblD pathogenic gene of B. cereus isolates. Mannitol egg yolk polymyxin agar was used as a selective isolation medium. Commercially available kits and boiling methods were used for DNA extraction, samples acquired from restaurants were examined for the presence of Hemolysin BL gene by polymerase chain reaction (PCR). Among all isolates, twenty-four of B. cereus isolates detected for HBL enterotoxin production by the discontinuous pattern on HBL sheep blood agar then confirmed by biochemical tests. More than 58.33 % of the isolate showed discontinuous hemolysis pattern on HBl blood agar and 29.16% of the samples were shown positive for hblD gene that can cause diarrhea with the size of 807bp on gel. This study demonstrated that RTE meat curry was a potential source for entero-toxigenic B. cereus and the presence of the hblD toxin genes for the HBL complex in the isolates tested were highly associated. Therefore, these meat curry isolates should be regarded as potential toxin producers.
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马来西亚餐馆咖喱肉中蜡样芽孢杆菌检测HBLD毒素基因
蜡样芽孢杆菌是一种普遍存在的食源性病原体,可引起食物中毒,导致感染,食物中毒有呕吐和腹泻两大类型。富含蛋白质的食物,如肉类,与蜡样芽孢杆菌引起的食源性腹泻暴发有关。本研究的目的是从马来西亚餐馆的即食咖喱肉中分离鉴定蜡样芽孢杆菌,并检测蜡样芽孢杆菌hblD致病基因。以甘露醇蛋黄多粘菌琼脂为选择性分离培养基。采用市售试剂盒和煮沸法提取DNA,采用聚合酶链反应(PCR)检测餐馆样品中溶血素BL基因的存在。在所有分离株中,有24株蜡样芽孢杆菌通过HBL羊血琼脂上的不连续模式检测出HBL肠毒素产生,并经生化试验证实。超过58.33%的分离物在HBl血琼脂上显示不连续溶血模式,29.16%的分离物显示hblD基因阳性,该基因可引起凝胶上大小为807bp的腹泻。本研究表明,RTE咖喱肉是产肠毒素蜡样芽孢杆菌的潜在来源,并且在测试的分离物中hblD毒素基因的存在与HBL复合物高度相关。因此,这些肉咖喱分离株应被视为潜在的毒素生产者。
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