Das Sanjoy, Surendran P K, Thampuran Nirmala K
Central Institute of Fisheries Technology, Cochin, India.
Indian J Med Res. 2009 Mar;129(3):316-20.
BACKGROUND & OBJECTIVE: Bacillus cereus is an important enterotoxigenic food borne pathogen. The present study was undertaken to assess the occurrence of B. cereus in tropical fish and evaluation of virulent gene specific PCR for differentiation of diarrhoeal enterotoxin producing isolates of B. cereus from non enterotoxigenic isolates.
Selective plating on polymixin-pyruvate-egg yolk-mannitol-bromocresol purple agar (PEMPA) was used for isolation of B. cereus from finfish, prawn and clams. Enterotoxin producing ability of all 42 isolates obtained from the samples was judged by reverse passive latex agglutination (RPLA) test and the presence of different virulent genes i.e. hbla, bceT and entFM was screened by PCR.
B. cereus and enterotoxigenic B. cereus were found to be in 36.7 and 29.41 per cent of fish samples, respectively. All the diarrhoeal enterotoxin producing isolates showed the presence of hbla gene, but hbla gene was not present in any of the non-enterotoxigenic isolates tested in this study.
INTERPRETATION & CONCLUSION: Our findings indicated that hbla gene specific PCR can be employed for differentiation of enterotoxigenic B. cereus isolates from non-enterotoxigenic isolates.
蜡样芽孢杆菌是一种重要的产肠毒素食源性病原体。本研究旨在评估热带鱼中蜡样芽孢杆菌的存在情况,并评价用于区分产腹泻性肠毒素的蜡样芽孢杆菌分离株与非产肠毒素分离株的毒力基因特异性聚合酶链反应(PCR)。
采用在多粘菌素 - 丙酮酸 - 蛋黄 - 甘露醇 - 溴甲酚紫琼脂(PEMPA)上进行选择性平板培养的方法,从鳍鱼、对虾和蛤中分离蜡样芽孢杆菌。通过反向被动乳胶凝集试验(RPLA)判断从样本中获得的所有42株分离株的产肠毒素能力,并通过PCR筛选不同毒力基因即hbla、bceT和entFM的存在情况。
蜡样芽孢杆菌和产肠毒素蜡样芽孢杆菌分别在36.7%和29.41%的鱼类样本中被发现。所有产腹泻性肠毒素的分离株均显示存在hbla基因,但在本研究中测试的任何非产肠毒素分离株中均未发现hbla基因。
我们的研究结果表明,hbla基因特异性PCR可用于区分产肠毒素蜡样芽孢杆菌分离株与非产肠毒素分离株。