Random amplified polymorphic DNA and restriction enzyme analysis of PCR amplified rDNA in taxonomy: two identification techniques for food-borne yeasts.

M M Baleiras Couto, J T Vogels, H Hofstra, J H Huis in't Veld, J M van der Vossen
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引用次数: 69

Abstract

The random amplified polymorphic DNA (RAPD) assay and the restriction enzyme analysis of PCR amplified rDNA are compared for the identification of the common spoilage yeasts Zygosaccharomyces bailii, Z. rouxii, Saccharomyces cerevisiae, Candida valida and C. lipolytica. Both techniques proved to be adequate tools for yeast identification. Since the RAPD does provide less stable patterns than restriction enzyme analysis of PCR amplified rDNA, and a large amount of data had to be compared without data reduction, Principal Component Analysis (PCA) was applied successfully for clustering the RAPD patterns. The success of PCA is highly influenced by the primer used in RAPD and the amount of reference samples. A large amount of reference samples improves the performance of clustering in PCA. The primer of choice was shown to be important with respect to the discriminatory power of the RAPD method. Some primers used enabled discrimination on the subspecies level. The results collected with both typing methods justify the conclusion that the present typing system can be applied for taxonomical purposes.

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随机扩增多态性DNA和PCR扩增rDNA的限制性内切酶分析:食源性酵母的两种鉴定技术。
比较了随机扩增多态性DNA (RAPD)法和PCR扩增rDNA限制性内切酶法对常见腐败酵母菌(Zygosaccharomyces bailii, Z. rouxii, Saccharomyces cerevisiae, Candida valida和C. lipolytica)的鉴定。这两种技术都被证明是酵母鉴定的适当工具。由于RAPD提供的模式不像PCR扩增的rDNA的限制性内切酶分析那样稳定,并且需要比较大量的数据而不进行数据缩减,因此应用主成分分析(PCA)成功地对RAPD模式进行聚类。主成分分析的成功与否很大程度上取决于RAPD中使用的引物和参考样品的数量。大量的参考样本提高了主成分分析的聚类性能。结果表明,引物的选择对RAPD方法的鉴别能力很重要。一些引物在亚种水平上进行了区分。两种分型方法收集的结果证明,该分型系统可用于分类学目的。
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