一种适用于聚合酶链反应的常见念珠菌基因组DNA提取方法的研究

A new PCR-suitable approach for DNA extraction from common pathogenic Candida

目的介绍一种简单、快速的真菌DNA提取方法,提高真菌DNA提取效率,减少毒性和污染性,以适应临床研究需要。方法同时用溶细胞酶结合Biospin真菌基因组DNA提取试剂盒和十六烷基三甲基溴化铵(CTAB)法,提取白色念珠菌、热带念珠菌、克柔念珠菌和净平滑念珠菌基因组DNA,用A260/A280的比值检测DNA的纯度并计算质量浓度,同时行聚合酶链反应(PCR)以评价其可靠性。结果溶细胞酶结合Biospin真菌基因组DNA提取试剂成功提取所用真菌基因组DNA,其纯度及质量浓度能满足PCR反应的要求。结论用溶细胞酶结合Biospin真菌基因组DNA提取试剂提取DNA,简单、快速、高效,可用于PCR反应。

Objective To introduce an approach of DNA extraction from fungus that is simple and quick, with high extraction efficiency, lower toxicity and less contamination, and therefore meets the need of clinical research. Methods Combination of lyticase with Biospin Fungus Genomic DNA Extraction Kit was used to extract DNA from pathogenic Candida, including Candida albicans Candida tropicalis, Candida Krusei and Candida Parapsilosis, compared with the same procedure using CTAB. The DNA purity was examined per ratio of A260/A280 and it concentration was calculated. The efficiency of this procedure was estimated using PCR. Results The genomic DNA of all tested fungi was extracted successfully with combined use of lyticase and Biospin Fungus Genomic DNA Extraction Kit, yielding satisfactory purity and concentration suitable for PCR. Conclusion Extraction of genomic DNA from Candida with combined use of lyticase and Biospin Fungus Genomic DNA Extraction Kit can be a convenient, rapid and efficient approach for purpose of PCR.