聚合酶链反应技术检测申克孢子丝菌的实验研究

目的研究申克孢子丝菌的种特异性引物聚合酶链反应鉴定方法,从而为临床孢子丝菌病的分子诊断奠定基础。方法采用根据申克孢子丝菌几丁质合成酶基因1序列设计合成的一对寡核苷酸引物,分别对26株申克孢子丝菌(包括1株标准株,7株实验室保存株,18株临床分离株)及6种6株普通真菌(分属于3个菌属)的基因组DNA进行PCR扩增。结果扩增产物选择性的从26株申克孢子丝菌基因组DNA中获得,而对念珠菌属、毛癣菌属、小孢子菌属的6株普通真菌基因组DNA均为阴性。结论采用以申克孢子丝菌几丁质合成酶基因1序列为基础的聚合酶链反应法鉴定申克孢子丝菌特异、简便、快捷,可用于临床诊断。

Laboratory Application of PCR for the Identification of Sporothrix Scheckii

Objective To study a rapid method to detect and identify Sporothrix schenckii by using Species-specific oligonucleotide primer PCR techniques, and lay the fundation of molecular diagnosis for sperotrichesis. Methods The Species - specific oligonucleotide primer pair S2-R2 was used in this study. It was designed from nucleotide sequences of chitin synthase 1 gene in Sporothrix schenckii. 26 strains of Sporothrix schenckii ,6 strains of clinical common fungi were amplified by PCR. 26 strains of Sporothrix schenckii include 1 standard strain,7 strains of laboratory reserved and 18 strains of clinical isolated. Results All strains of Sporothrix schenckii showed a specific fragment of 318bp with the species-specific primer pair. Using the same primer pairs, the others species had no specific amplification. Conclusion This method is specific, rapid and simple for identifying Sporothrix schenckii and could be used for clinical molecular diagnosis.