关键词: 申克孢子丝茵, 基因组DNA提取, 聚合酶链反应, DNA引物, 钙调蛋白基因

Abstract:

Objectives A modified method for extracting DNA from Sporothrix. To study a rapid method to detect and identify sporothrix schenckii by using Species-specific oligonucleotide primer PCR techniques,and lay the foundation of molecular diagnosis for sporotrichosis. Methods Viscozyme L enzyme was used to replace of the traditional method. The species-specific oligonucleotide primer pair was used in this study. It was designed from nucleotide sequences of calmodulin gene in Sporothrix schenckii. 52 strains of Sporothrix schenckii and 6 strains of common fungus were amplified by PCR. Results Genomic DNA of Sporothrix schenckii were extracted with the amended (CTAB)method. All strains of Sporothrix schenckii showed a specific fragment of about 430bp with the species-specific primer pair. Using the same primer pair,the other species had no specific amplification. Conclusions Compared with the traditional method,higher yield and purity of genomic DNA were obtained with less contamination. The result indicted that this was a simple and highly efficient method. This method was specific,sensitive,reliable,rapid and simple for identifying Sporothrix schenckii and could be used for clinical molecular diagnosis.

Key words: Sporothrix schenckii, genomic DNA extraction, PCR, DNA primers, calmodulin gene