玻璃珠-盐析法提取常见致病真菌DAN的研究

Glass Beads－ Salting out Procedure for Extracting DNA from Common Pathogenic Fungi

目的 进一步提高提取 DNA的效率并减少污染和毒性，以适应临床 PCR检测的需要，建立玻璃珠－盐析法快速提取常见致病真菌 DNA。方法 分别采用玻璃珠－盐析法和经典的酶解－酚氯仿法比较从白念珠菌、乳酒念珠菌、克柔念珠菌、季也蒙念珠菌、新生隐球菌、烟曲霉、黄曲霉和马内菲青霉等 8种常见致病真菌中提取 DNA，同时行 PCR以评价其可靠性。 结果 本方法从 8种常见致病真菌均提取浓度较高且较完整的 DNA，用于 PCR也获得良好结果。结论 玻璃珠－盐析法提取致病酵母菌和丝状真菌安全、快速且有效。

Objective To develop glass－ beads and salting－ out procedure for extracting DNA from common pathogenic fungi. Methods Glass beads－ salting out procedure was used to isolated DNA from 8 pathogenic fungi, including Candida albicans, Candida kefyr, Candida krusei, Candida guilliermondii, Cryptococcus neoformans, Aspergillus fumigatus, Aspergillus flavus and Penicillium marneffei, and compared with conventional phenol－ chloroform extraction method. The efficiency of this procedure was estimated by using PCR. Results For all the eight organisms, the rapid extraction procedure resulted in high yield and intact DNA. Conclusion The disruption of fungal cells by glass beads－ salting out procedure is a safe, rapid, and efficient method for extraction of genomic DNA from medically important yeasts and filamentous fungi.