申克孢子丝菌随机扩增DNA多态性研究

Characterization of Sporothrix schenckii by Random Amplification of Polymorp hic DNA Assay

目的了解申克孢子丝菌的基因学特征,研究DNA分型与菌种来源和临床表现的关系。方法以随机扩增DNA多态性方法对来源于不同地区及不同临床类型的24株申克孢子丝菌进行DNA多态性分析。结果①筛选出3个具有较好扩增片段的引物,即引物OPAA11:5'-ACCCGACCTG-3',OPD18:5'-GAGAGCCAAC-3',OPB07:5'-GGTGACGCAG-3'。②24株菌的DNA带型不完全相同,具一定遗传变异性。③不同菌株同一引物扩增均见一共有DNA片段。④播散型DNA带型与皮肤淋巴管型不同。结论随机扩增DNA多态性方法可用于申克孢子丝菌的DNA分型。其带型与菌株的地区来源和临床表现有关。

Objective To investigate the DNA polymorphism of S.schenckii and to find the ralationship between DNA patterns,geographic areas,and clinical manifestations.Methods The total DNA was extracted with hexadecyltrimethylammonium b romide.Random amplification of polymorphic DNA(RAPD)assay was used to study DNA typing in 24strains of S.schenckii which were collected from different areas and from different clinical types.Results①Of seven random primers used,three p rimers(OPAA11?OPD18and OPB07)gave good reactions,the sequences of whi ch were 5' -ACCCGACCTG -3',5' -GAGAGCCAAC -3',5' -GGT-GACGCAG -3' ,respectively.②The RAPD patterns of the 24isolates w ere not identical,showing certain degrees of hereditary variation.③Different isolates showed a common c onserved DNA band with the same primer.④There were different genotypes betw een different clinical types.Conclusion According to RAPD typing of S.schenckii,the DNA band type is related to its geographic origin and clinical manife station.