【摘要】 目的利用探针与DNA印迹法对申克孢子丝菌进行种内分型,探讨其基因型特征与菌种来源及临床表现的关系。方法CTAB法提取来源于不同地区31株孢子丝菌临床分离株及1株标准株的基因组DNA,以真菌通用引物ITS4、NS5扩增标准株的rDNA序列作为探针,与经限制性内切酶ApaI酶切后的基因组DNA进行印迹杂交。结果杂交后形成多种清晰而稳定的带型,根据带型将31株孢子丝菌分为15种基因型(A-O型),其中A、B、C三型占51.61%。结论探针与DNA印迹法是孢子丝菌种内分型较为敏感而可靠的方法,该方法所分基因型的不同与菌种的地区来源及临床表现有一定的相关性。更多还原

【Abstract】 Objectives To identify genotypes of 31 Sporothrix schenckii (S.schenckii) strains by Southern blotting and to explore the relationship between genotypes and geographic distributions and clinical manifestations.Methods Total DNA was extracted by cetyltriethyl ammonium bromide (CTAB).The polymorphisms were detected by hybridization of ApaⅠ-digested S.schenckii genomic DNA with a probe amplified from the small-subunit rDNA and adjacent internal transcribed spacer (ITS) regions.The band patterns manifested by Southern blotting were employed to investigate genotypes of 31 strains of S.schenckii collected from five different areas in China.Results Of 31 strains of S.schenckii,15 individual patterns (DNA Type A-O) were recognized.Type A to C accounted for 51.61% of all strains.Conclusion The Southern blotting provides a highly sensitive and reliable means for DNA typing of S.schenckii.It is also found that there is an obvious correlation between DNA patterns and different geographic distribution and clinical manifestations.更多还原