Feurer C, Irlinger F, Spinnler H E, Glaser P, Vallaeys T
Laboratoire de Génie et Microbiologie des Procédés alimentaires, INRA, Thiverval Grignon, France.
J Appl Microbiol. 2004;97(3):546-56. doi: 10.1111/j.1365-2672.2004.02333.x.
The diversity of the surface flora of two French red-smear soft cheeses was examined by cultivation-dependent and cultivation-independent methods to assess their composition and to evaluate the accuracy of both approaches.
Culture-independent methods used involved 16S ribosomal DNA gene cloning and sequencing and single-strand conformation polymorphism analysis (SSCP). The culture-dependent method used involved direct culture and macroscopic observation, polymerase chain reaction of the 16S rRNA gene from DNA extracted from single colonies followed by complete sequencing of the gene. Only few species were recovered by both approaches either in the pasteurized and the farmer cheese. A large diversity of isolates or 16S rDNA sequences related to marine bacteria was identified at the surface of both cheeses.
The results indicated that all three techniques were informative and complementary to allow a more accurate representativeness of the cheese surface biodiversity.
Cultivation and molecular methods have to be combined in order to obtain an extended view of the bacterial populations of complex ecosystems.
通过依赖培养和不依赖培养的方法研究两种法国红抹软质奶酪表面菌群的多样性,以评估其组成并评价这两种方法的准确性。
所采用的不依赖培养的方法包括16S核糖体DNA基因克隆与测序以及单链构象多态性分析(SSCP)。所采用的依赖培养的方法包括直接培养和宏观观察,从单个菌落提取的DNA进行16S rRNA基因的聚合酶链反应,随后对该基因进行全序列测定。在巴氏杀菌奶酪和农家奶酪中,两种方法仅回收了少数几种菌种。在两种奶酪的表面均鉴定出了与海洋细菌相关的多种分离株或16S rDNA序列。
结果表明,所有这三种技术都提供了信息且相互补充,从而能够更准确地呈现奶酪表面生物多样性。
必须将培养方法和分子方法结合起来,以便更全面地了解复杂生态系统中的细菌种群。