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目的 为了判断临床和实验室标本中是否存在病原性真菌。方法 设计了一个以热启动聚合酶链反应 (PCR)为基础的实验方法 ,以一段真菌特异性DNA序列作为通用引物进行检测。引物序列为 :①AACTTAAAGGAATTGACGGAAG ;②GCATCACAGACCTGTTATTGCCTC。结果 该法能在 3小时之内对所用的全部 2 3种共 42株重要病原性真菌及经培养证实有真菌的 2 2份临床标本成功地扩增出一条310bp的DNA片段 ,但对其它微生物和人体细胞均未扩增出类似片段。该法敏感性高 ,特异性强。结论 采用通用性强的引物系统配合特异性高的热启动PCR技术检测临床和实验室标本中是否存在病原性真菌的方法有重要的应用潜力。
Purpose To determine the presence of pathogenic fungi in clinical and laboratory specimens. Methods A novel thermostable polymerase chain reaction (PCR) -based assay was designed to detect a fungal-specific DNA sequence as a universal primer. Primer sequences: ① AACTTAAAGGAATTGACGGAAG; ② GCATCACAGACCTGTTATTGCCTC. Results The method was able to successfully amplify a 310 bp DNA fragment of all 2 3 42 pathogenic fungi and 2 2 clinically proven fungi used within 3 hours. However, for other microorganisms And human cells did not amplify a similar fragment. The method is highly sensitive and specific. Conclusion The method of using universal primer system and high specificity hot-start PCR to detect the presence of pathogenic fungi in clinical and laboratory samples has important potential.