PCR检测部分病原真菌的实验研究

为探讨和评价ＰＣＲ技术在检测和鉴定病原真菌方面的作用，重点是引物的选择。首先比较了来自ｒＤＮＡ基因序列的通用引物的通用性，并用３对种特异性引物进行２次套式扩增。结果表明３对通用引物均能扩增出预期的靶ＤＮＡ带，平均大小分别为６２０ｂｐ，４８５ｂｐ和５８０ｂｐ；３对种特异性引物分别扩增出白念珠菌、新生隐球菌和烟曲霉ＤＮＡ，并提高了敏感性。说明用通用引物和种特异引物进行ＰＣＲ扩增可以用于病原真菌的检测和

Study on the detection and identification of pathogenic fungi by PCR

To study and evaluate the application of PCR technique on the detection and identification of pathogenic fungi for the disgnosis of systemicmycoses. 3 pairs of universal fungal PCR primers derived from the rDNA were compared in universality and stability, and identification of the fungus was achieved by a second series of nested amplification with 3 pairs of species specific primers. 3 pairs of universal primers were capable of amplifying DNA from 30 strains of medically important fungi, and the median sizes of DNAs amplified were 620bp, 485bp and 580bp, respectively. C. albicans, C. neoformans and A. fumigatus specific forward primers were used in combination with the universal reverse primer. Using these primer- pairs only homologous DNA was amplified. The desribed PCR assay allows for the highly sensitive and specific detection(universal PCR) and identification(species- specific) of pathogenic fungi.