摘要
目的 :应用随机扩增多态性 DNA( RAPD)技术对临床上常见的酵母菌进行分析 ,以选择合适的引物和实验条件对酵母菌进行鉴定。方法 :选用四条长度为 10个碱基的随机引物分别对酵母菌、丝状真菌及细菌进行扩增 ,并对扩增结果进行相似性分析。结果 :所选三条引物可在种的水平上将所试菌株有效地区分开来 ,但同一引物对不同菌种的扩增结果存在明显的差异。在扩增同种内不同菌株时 ,各菌株间也存在着一定的差异。采用不同引物进行扩增时种内变异的程度有所不同。除个别菌株外 ,扩增所得指纹图型的种类差异远远小于不同的菌种间的差异 ,因而可将不同种的酵母菌有效地区分开来。结论 :RAPD技术可用于酵母菌的种间鉴定。其中引物及实验条件的选择尤为重要 ,同时 。
Objective:To analysis DNA polymorphism in different species and strains of the medically important yeasts by random amplified polymorphic DNA (RAPD). Methods:By using four short oligonucleotide primers (10 mers) with arbitrarily chosen sequences in the polymerase chain reaction, we detected 13 different yeasts、 2 other fungal species and 2 bacteria. Similarity analysis was also employed. Results:The number and size of the amplification products were characteristic for each species. Interspecies variation was observed within the genus yeasts, and species colud be clearly distinguished by their RAPD profiles. The RAPD profile derived from different primers had distinct diversity. By using same primer, intraspecies DNA length polymorphisms were seen for RAPD profiles and similarity analysis from different species. Especially, the intraspecies polymorphism from Crytococcus neoformans was very obvious. By using different primers, the intraspecies variation of RAPD profiles was different. Except few strains, using primer Ⅰ、Ⅱ、Ⅳ, the variation among profiles obtained from different strains of the same species was far less than the variation observed among different species of yeasts. So different yeast species could be distinguished by their RAPD profiles.Conclusion:RAPD can be used in the interspecies detection of Yeasts. It is very important to choose primers and experimental conditions. The reliability of detection would be improved by using combinations of oligonucleotide primers.
出处
《军医进修学院学报》
CAS
2000年第1期73-76,共4页
Academic Journal of Pla Postgraduate Medical School
