Genetic identification and detection of human pathogenic Rhizopus species, a major mucormycosis agent, by multiplex PCR based on internal transcribed spacer region of rRNA gene
Genetic identification and detection of human pathogenic Rhizopus species, a major mucormycosis agent, by multiplex PCR based on internal transcribed spacer region of rRNA gene
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DOI：
10.1016/j.jdermsci.2005.01.010
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发表时间：
2005-07-01
影响因子：
4.6
通讯作者：
Nishikawa, T
中科院分区：
文献类型：
--
作者：
Nagao, K;Ota, T;Nishikawa, T
背景：毛霉菌病是一种由毛霉目的真菌引起的侵袭性机会性感染。由于缺乏实验室检查，毛霉菌病的诊断是出了名的困难。再加上其快速的进展，以及虚弱的状态，病人谁合同的疾病，死亡率是非常高的目的：本研究的目的是遗传学鉴定的根霉，一个主要的毛霉菌病的代理人，通过rRNA基因的内转录间隔区（ITS）区域。PCR不仅与测试菌株和临床分离株，而且与皮肤毛霉菌病患者的临床样品。结果：多重PCR共鉴定出5种根霉，其中根霉（Rhizopus sp.）azygosporus只能与R.小孢子虫的基因序列中存在某些多态性。当这种多重PCR应用于从三个毛霉病患者（石蜡切片从所有和血清从一个病人）的临床样本，根霉DNA对应的分离的病原体进行了具体detected.Conclusion：虽然从临床样本的真菌DNA检测是一个严格的研究领域，这是第一次报告，以遗传学鉴定和检测人类毛霉病标本根霉属物种。这可能扩大这种多重PCR系统的可能性，不仅要确定分离的真菌，但也作为内脏毛霉菌病的筛选方法。(c)2005年日本皮肤病研究学会。由Elsevier爱尔兰有限公司出版。保留AR权利。
Background: Mucormycosis is an invasive opportunistic infection caused by fungi belonging to the order Mucorates. Due to the lack of laboratory tests, the diagnosis of mucormycosis is notoriously difficult. Added with its rapid progression as well as the debilitated state of the patients who contract the disease, mortality is extremely high.Objective: The goal of this study was to genetically identify human pathogenic Rhizopus species, a major mucormycosis agent, by the internal transcribed spacer (ITS) region of rRNA gene.Methods: Primers were designed to identify five Rhizopus species known to cause human disease by multiplex PCR. PCR was done not only with test strains and clinical isolates, but also with clinical samples from cutaneous mucormycosis patients. Sporangiospore morphology was observed by scanning electron microscopy to confirm the correlation of phenotypic and genotypic features.Results: Multiplex PCR identified five Rhizopus species including Rhizopus oryzae, where R. azygosporus could only be distinguished from R. microsporus by certain polymorphisms that were present in its sequence. When this multiplex PCR was applied to clinical samples from three mucormycosis patients (paraffin sections from all and sera from one patient), Rhizopus DNA corresponding to the isolated pathogens were specifically detected.Conclusion: While fungal DNA detection from clinical samples is a rigorously studied area, this is the first report to genetically identify and detect Rhizopus species from human mucormycosis specimens. This may expand the possibility of this multiplex PCR system not only to identify isolated fungi, but also as a screening method for visceral mucormycosis. (c) 2005 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. AR rights reserved.