Manzano Marisa, Cocolin Luca, Cantoni Carlo, Comi Giuseppe
Dipartimento di Scienze degli Alimenti, Facoltà di Agraria, Università degli Studi di Udine, Italy.
Int J Food Microbiol. 2003 Mar 25;81(3):249-54. doi: 10.1016/s0168-1605(02)00222-2.
A method was developed to differentiate between Bacillus cereus, Bacillus mycoides and Bacillus thuringiensis using the polymerase chain reaction combined with a restriction endonuclease (PCR-RE) technique. This fast and simple protocol, applied to pure culture strains, was developed using the gyrB DNA sequence, as previously proposed by other authors. Strains from international collections were used to optimize the method which was then applied to the identification of strains isolated from food samples. Amplifications were specific for the B. cereus group. Only Staphylococcus aureus gave the same size PCR product, but it was easily differentiated from strains in the B. cereus group by using restriction analysis, based on digestion with the RsaI, Sau3AI and EcoRI endonucleases. Specific amplifications and good differentiations were obtained using pure strains, suggesting the possibility of using the method described to identify the B. cereus group directly in food samples.
开发了一种使用聚合酶链反应结合限制性内切酶（PCR-RE）技术区分蜡样芽孢杆菌、蕈状芽孢杆菌和苏云金芽孢杆菌的方法。该快速简便的方案应用于纯培养菌株，是根据其他作者先前提出的方法，利用gyrB DNA序列开发的。使用来自国际菌种保藏中心的菌株优化该方法，然后将其应用于从食品样品中分离的菌株的鉴定。扩增对蜡样芽孢杆菌群具有特异性。只有金黄色葡萄球菌产生相同大小的PCR产物，但通过使用基于RsaI、Sau3AI和EcoRI内切酶消化的限制性分析，很容易将其与蜡样芽孢杆菌群中的菌株区分开来。使用纯菌株获得了特异性扩增和良好的区分效果，这表明有可能使用所述方法直接在食品样品中鉴定蜡样芽孢杆菌群。
