Bu Rong, Sathiapalan Rajeev K, Ibrahim Muna M, Al-Mohsen Ibrahim, Almodavar Edna, Gutierrez Marina I, Bhatia Kishor
King Fahad National Center for Children's Cancer and Research, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia 2King Fasial Specialist Hospital and Research Center, Riyadh, Saudi Arabia 3International Network for Cancer Treatment and Research, Brussels, Belgium.
J Med Microbiol. 2005 Mar;54(Pt 3):243-248. doi: 10.1099/jmm.0.45856-0.
Invasive fungal pathogens, especially in immunocompromised hosts, can result in life-threatening infections. Current laboratory/radiological methods for fungal identification are time-consuming and lack sensitivity and specificity. A monochrome, multiplex, real-time PCR assay for the identification and quantification of Candida albicans, Candida krusei, Candida tropicalis, Aspergillus flavus and Aspergillus fumigatus is described here. Detection of each of these fungi was specific and demonstrated 100 % concordance with biochemical/culture identification in all 60 isolates tested. Samples from 16 febrile neutropenic patients with haematological malignancies were also analysed and the utility of the assay in clinical samples was reconfirmed without false-negative results. The sensitivity of this assay was 0.1 pg fungal genomic DNA, corresponding to three cells, for C. albicans, C. krusei, C. tropicalis and A. flavus, and 0.01 pg fungal genomic DNA, i.e. less than one cell, for A. fumigatus. The analysis allows a low-cost, simple, rapid and sensitive alternative for clinical identification and quantification of these five common fungal species.
侵袭性真菌病原体，尤其是在免疫功能低下的宿主中，可导致危及生命的感染。目前用于真菌鉴定的实验室/放射学方法耗时且缺乏敏感性和特异性。本文描述了一种用于白色念珠菌、克鲁斯念珠菌、热带念珠菌、黄曲霉和烟曲霉鉴定及定量的单色、多重、实时PCR检测方法。对这些真菌中的每一种的检测都是特异的，并且在所有60株测试菌株中与生化/培养鉴定显示出100%的一致性。还分析了16例血液系统恶性肿瘤发热性中性粒细胞减少患者的样本，再次证实了该检测方法在临床样本中的实用性，且无假阴性结果。该检测方法对白色念珠菌、克鲁斯念珠菌、热带念珠菌和黄曲霉的敏感性为0.1 pg真菌基因组DNA，相当于三个细胞，对烟曲霉的敏感性为0.01 pg真菌基因组DNA，即少于一个细胞。该分析为这五种常见真菌物种的临床鉴定和定量提供了一种低成本、简单、快速且灵敏的替代方法。
