Specific Detection of Fusarium Species in Blood and Tissues by a PCR Technique
Specific Detection of Fusarium Species in Blood and Tissues by a PCR Technique
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利用 PCR 技术特异性检测血液和组织中的镰刀菌种类
DOI：
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发表时间：
1999
影响因子：
9.4
通讯作者：
C. de Bièvre
中科院分区：
文献类型：
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作者：
Francois;M. Huerre;Marie Ange Rouffault;C. de Bièvre
镰刀菌是一种机会性的医院病原菌，常引起致死性侵袭性真菌病。我们设计了一个引物对，通过聚合酶链反应（PCR）扩增了一段编码褐藻rRNA的基因片段。这些引物扩增出了主要镰刀菌属和vas感染性新宇宙孢子菌属的dna，但未扩增出11种重要医学真菌的dna。采用溴化乙啶染色法对茄枯菌DNA的检测下限为10 fg。为了测试该PCR系统检测组织中镰刀菌DNA的能力，我们建立了播散性镰刀菌病的小鼠模型。利用PCR技术，我们在小鼠组织和人血中检测到镰刀菌DNA。利用随机扩增多态性DNA （RAPD）对番茄枯萎菌、moniliform镰刀菌和尖孢镰刀菌进行检测。对RAPD分析产生的条带进行纯化、克隆和测序。这些信息被用来设计引物对，选择性地扩增一个或几个镰刀菌。该方法可用于培养和临床样品镰刀菌的快速检测和鉴定。
ABSTRACT Fusarium species are opportunistic nosocomial pathogens that often cause fatal invasive mycoses. We designed a primer pair that amplifies by PCR a fragment of a gene coding for the rRNA ofFusarium species. The DNAs of the main Fusariumspecies and Neocosmospora vasinfecta but not the DNAs from 11 medically important fungi were amplified by these primers. The lower limit of detection of the PCR system was 10 fg of Fusarium solani DNA by ethidium bromide staining. To test the ability of this PCR system to detect Fusarium DNA in tissues, we developed a mouse model of disseminated fusariosis. Using the PCR, we detected Fusarium DNA in mouse tissues and in spiked human blood. Furthermore, F. solani, Fusarium moniliforme, and Fusarium oxysporum were testing by random amplified polymorphic DNA (RAPD) analysis. The bands produced by RAPD analysis were purified, cloned, and sequenced. The information was used to design primer pairs that selectively amplified one or severalFusarium species. The method developed may be useful for the rapid detection and identification of Fusarium species both from culture and from clinical samples.
通过聚合酶链反应进行死后镰刀菌全眼炎诊断。
影响因子：
4.2
作者：
Alexandrakis,G;Sears,M;Gloor,P
通讯作者：
Gloor,P