Fortina M G, Ricci G, Borgo F, Manachini P L
Industrial Microbiology Section, Department of Food Science and Microbiology, University of Milan, Milan, Italy.
Lett Appl Microbiol. 2007 Apr;44(4):443-6. doi: 10.1111/j.1472-765X.2006.02082.x.
To develop a species-specific PCR assay with primers targeted to 16S rRNA gene for the identification of Enterococcus italicus, a new species of Enterococcus, involved in the production of Italian cheeses.
The type strain of E. italicus (DSM 15952(T) - 16S rRNA gene accession no. AJ582753) and other strains of the species were subjected to a rapid identification by PCR using primer pairs located within the 16S rRNA gene. A species-specific PCR product of approximately 323 bp was obtained after amplification of all E. italicus strains tested. The specificity of the primers was validated with representatives of the most closely related genera and species and a number of other bacterial species. In addition, the technique enabled the recognition of E. italicus from cheeses.
The protocol was highly efficient and sensitive, enabling the identification of E. italicus from cheeses.
The species-specific PCR offers a reliable and rapid alternative to conventional phenotypic methods for the identification of E. italicus within the heterogeneous genus Enterococcus.
开发一种针对16S rRNA基因的物种特异性聚合酶链反应(PCR)检测方法,用于鉴定意大利肠球菌,这是一种参与意大利奶酪生产的新型肠球菌。
使用位于16S rRNA基因内的引物对,通过PCR对意大利肠球菌模式菌株(DSM 15952(T) - 16S rRNA基因登录号AJ582753)及该物种的其他菌株进行快速鉴定。对所有测试的意大利肠球菌菌株进行扩增后,获得了约323 bp的物种特异性PCR产物。用最密切相关属和种的代表以及许多其他细菌物种验证了引物的特异性。此外,该技术能够从奶酪中识别出意大利肠球菌。
该方法高效且灵敏,能够从奶酪中鉴定出意大利肠球菌。
物种特异性PCR为在异质性肠球菌属中鉴定意大利肠球菌提供了一种可靠且快速的传统表型方法的替代方法。