Klein Günter
Bundesinstitut für Risikobewertung, Berlin, und Zentrum für Lebensmittelwissenschaft, Tierärztliche Hochschule, Hannover.
Berl Munch Tierarztl Wochenschr. 2003 Nov-Dec;116(11-12):510-6.
The aim of molecular methods in food microbiology is the identification or strain specific differentiation of microorganisms. Identification methods include besides taxonomic purposes also the detection of virulence genes or resistance markers. Strain specific differentiation is used for epidemiological investigations or the quality control of technologically used bacteria. Probiotic strains of the Lactobacillus acidophilus-group were investigated with different molecular methods: for identification proteinfingerprinting and RAPD-PCR (Random Amplified Polymorphic DNA-PCR) were applied, for strain specific differentiation pulsed field gel electrophoresis (PFGE) and again RAPD-PCR were used. The molecular methods applied should be chosen depending on the objective (identification, differentiation) and with respect to the organism to be tested. In case of probiotic lactic acid bacteria like the L. acidophilus-group proteinfingerprinting has proved to be successful for identification and with some limitation also RAPD-PCR. For differentiation PFGE is suitable as well as RAPD-PCR. Those methods differ substantially in their work load and in personal requirements, and show different power of discrimination and reproducibility within and between laboratories. This should be considered while choosing the appropriate method.
食品微生物学中分子方法的目的是对微生物进行鉴定或菌株特异性区分。除分类学目的外,鉴定方法还包括检测毒力基因或抗性标记。菌株特异性区分用于流行病学调查或对工业用细菌的质量控制。采用不同的分子方法对嗜酸乳杆菌属的益生菌株进行了研究:用于鉴定的有蛋白质指纹图谱法和随机扩增多态性DNA-PCR(RAPD-PCR),用于菌株特异性区分的有脉冲场凝胶电泳(PFGE)以及再次使用的RAPD-PCR。应根据目标(鉴定、区分)并针对待检测的生物体选择所应用的分子方法。对于嗜酸乳杆菌属这类益生菌,蛋白质指纹图谱法已被证明在鉴定方面是成功的,RAPD-PCR在一定程度上也适用。对于区分,PFGE和RAPD-PCR都适用。这些方法在工作量和人员要求方面有很大差异,并且在实验室内部和实验室之间表现出不同的鉴别力和重现性。在选择合适的方法时应考虑到这一点。