Venkateswaran K, Murakoshi A, Satake M
Nippon Suisan Kaisha, Ltd., Central Research Laboratory, Tokyo, Japan.
Appl Environ Microbiol. 1996 Jul;62(7):2236-43. doi: 10.1128/aem.62.7.2236-2243.1996.
Three commercially available kits that were supplemented with substrates for enzyme reactions were evaluated to determine their abilities to detect coliforms and fecal coliforms in foods. Japanese and U.S. Food and Drug Administration standard methods, as well as two agar plate methods, were compared with the three commercial kits. A total of 50 food samples from various retailers were examined. The levels of detection of coliforms were high with the commercial kits (78 to 98%) compared with the levels of detection with the standard methods (80 to 83%) and the agar plate methods (56 to 83%). Among the kits tested, the Colilert kit had highest level of recovery of coliforms (98%), and the level of recovery of Escherichia coli as determined by beta-glucuronidase activity with the Colilert kit (83%) was comparable to the level of recovery obtained by the U.S. Food and Drug Administration method (87%). Isolation of E. coli on the basis of the beta-glucuronidase enzyme reaction was found to be good. Levine's eosine methylene blue agar, which has been widely used in various laboratories to isolate E. coli was compared with 4-methylumbelliferyl-beta-D-glucuronide (MUG)-supplemented agar for isolation of E. coli. Only 47% of the E. coli was detected when eosine methylene blue agar was used; however, when violet red bile (VRB)-MUG agar was used, the E. coli detection rate was twice as high. Of the 200 E. coli strains isolated, only 2 were found to be MUG negative, and the gene responsible for beta-glucuronidase activity (uidA gene) was detected by the PCR method in these 2 strains. Of the 90 false-positive strains isolated that exhibited various E. coli characteristic features, only 2 non-E.coli strains hydrolyzed MUG and produced fluorescent substrate in VRB-MUG agar. However, the PCR did not amplify uidA gene products in these VRB-MUG fluorescence-positive strains.
对三种补充了酶反应底物的市售试剂盒进行了评估,以确定它们检测食品中大肠菌群和粪大肠菌群的能力。将日本和美国食品药品监督管理局的标准方法以及两种琼脂平板法与这三种商用试剂盒进行了比较。共检查了来自不同零售商的50份食品样本。与标准方法(80%至83%)和琼脂平板法(56%至83%)的检测水平相比,商用试剂盒对大肠菌群的检测水平较高(78%至98%)。在所测试的试剂盒中,Colilert试剂盒对大肠菌群的回收率最高(98%),通过Colilert试剂盒的β-葡萄糖醛酸酶活性测定的大肠杆菌回收率(83%)与美国食品药品监督管理局方法获得的回收率(87%)相当。基于β-葡萄糖醛酸酶反应分离大肠杆菌的效果良好。将在各个实验室广泛用于分离大肠杆菌的莱文伊红美蓝琼脂与补充了4-甲基伞形酮-β-D-葡萄糖醛酸(MUG)的琼脂用于分离大肠杆菌的情况进行了比较。使用伊红美蓝琼脂时,仅检测到47%的大肠杆菌;然而,使用紫红胆盐(VRB)-MUG琼脂时,大肠杆菌的检测率高出一倍。在分离出的200株大肠杆菌菌株中,仅发现2株为MUG阴性,并且通过PCR方法在这2株菌株中检测到了负责β-葡萄糖醛酸酶活性的基因(uidA基因)。在分离出的90株表现出各种大肠杆菌特征的假阳性菌株中,只有2株非大肠杆菌菌株在VRB-MUG琼脂中水解MUG并产生荧光底物。然而,PCR未在这些VRB-MUG荧光阳性菌株中扩增出uidA基因产物。