Lugert R, Schettler C, Gross U
National Reference Center for Systemic Mycoses, Institute of Medical Microbiology, University of Göttingen, Germany.
Mycoses. 2006 Jul;49(4):298-304. doi: 10.1111/j.1439-0507.2006.01255.x.
Although a large number of different PCR protocols for the detection of fungal DNA from clinical samples have been described, a generally recognised standardisation has not yet been developed. In a first step, we compared six different methods to isolate DNA under in vitro conditions from Aspergillus fumigatus, Candida albicans and Saccharomyces cerevisiae with respect to efficiency and expenditure of time. To this end, methods were tested that are based on both mechanical and enzymatic/thermic lysis. Thereby, enzymatic/thermic lysis were shown to be superior to mechanical lysis, although these methods of DNA isolation were more time consuming. The subsequent comparison of three different PCR protocols showed real-time PCR to be the most sensitive method.
虽然已经描述了大量用于从临床样本中检测真菌DNA的不同PCR方案，但尚未形成普遍认可的标准化方法。第一步，我们比较了六种不同的方法，在体外条件下从烟曲霉、白色念珠菌和酿酒酵母中分离DNA，比较其效率和时间消耗。为此，测试了基于机械裂解和酶解/热裂解的方法。结果表明，酶解/热裂解优于机械裂解，尽管这些DNA分离方法耗时更长。随后对三种不同PCR方案的比较表明，实时PCR是最灵敏的方法。
