Coppola S, Blaiotta G, Ercolini D, Moschetti G
Dipartimento di Scienza degli Alimenti, Sezione di Microbiologia Agraria, Alimentare ed Ambientale e di Igiene, Università degli Studi di Napoli 'Federico II', Portici, Italy.
J Appl Microbiol. 2001 Mar;90(3):414-20. doi: 10.1046/j.1365-2672.2001.01262.x.
The microbial community of different types of unripened Pasta Filata cheese was investigated by culture-independent methods with the aim of rapidly achieving knowledge about cheese microbiota and discriminating traditional and industrial cheeses.
The microbial DNA extracted directly from the samples was used as a template in PCR experiments to amplify the 16S-23S rDNA spacer region and the V3 region of the 16S rDNA. Conventional electrophoresis of the amplified spacers allowed known classes of these DNA fragments belonging to genera and species of lactic acid bacteria to be distinguished. Denaturing gradient gel electrophoresis analysis of V3 amplicons was supported by reference cultures of LAB used as markers.
Both molecular approaches furnished the expected information about microbial diversity and were quite valid for discriminating industrial, semi-artisanal or traditional cheeses, characterized by increasingly complex DNA profiles.
Both methods could be used for legal purposes when products obtained through prescribed manufacturing regulations are to be analysed.
采用非培养方法研究不同类型的未成熟丝状干酪的微生物群落，旨在快速了解干酪微生物群，并区分传统干酪和工业干酪。
直接从样品中提取的微生物DNA用作PCR实验的模板，以扩增16S - 23S rDNA间隔区和16S rDNA的V3区。扩增间隔区的常规电泳可区分属于乳酸菌属和种的这些DNA片段的已知类别。V3扩增子的变性梯度凝胶电泳分析得到用作标记的乳酸菌参考培养物的支持。
两种分子方法都提供了有关微生物多样性的预期信息，并且对于区分具有日益复杂DNA图谱特征的工业、半手工或传统干酪非常有效。
当要分析通过规定制造法规获得的产品时，这两种方法都可用于法律目的。
