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[使用分子遗传学方法检测和鉴定引起真菌病的丝状真菌]

[Detection and identification of filamentous fungi causing mycoses using molecular genetic methods].

作者信息

Bejdák Petr, Lengerová Martina, Paloušová Dita, Volfová Pavlína, Kocmanová Iva, Mayer Jiří, Ráčil Zdeněk

机构信息

Center of Molecular Biology and Gene Therapy, Department of Internal Medicine - Hematology and Oncology, University Hospital Brno and Masaryk University, Czech Republic.

出版信息

Klin Mikrobiol Infekc Lek. 2012 Aug;18(4):109-14.

Abstract

Methods of molecular genetics offer rapid and sensitive detection and identification of fungal pathogens. The currently used methods are based mainly on PCR. With regard to the ubiquitous presence of fungi, it is important to prevent contamination during the whole process, from sampling to laboratory analyses. Molecular genetic methods are not included among the EORTC/MSG criteria used for the diagnosis of invasive fungal diseases since interlaboratory standardization is still missing. Another reason is the use of different target genes for PCR. ITS sequences from rDNA clusters are recommended for DNA barcoding of fungi. The use of DNA sequencing for identification of fungi in clinical samples has certain limitations and interpretation of results could be problematic in some cases. DNA sequences are searched and compared in public databases on the Internet, the best known of them being the GenBank. However, more reliable data for identification of fungi are offered by specialized mycological databases.

摘要

分子遗传学方法能够快速、灵敏地检测和鉴定真菌病原体。目前使用的方法主要基于聚合酶链反应(PCR)。鉴于真菌无处不在,从采样到实验室分析的整个过程中防止污染至关重要。分子遗传学方法未被纳入欧洲癌症研究与治疗组织/侵袭性真菌感染协作组(EORTC/MSG)用于诊断侵袭性真菌病的标准中,因为实验室间的标准化仍然缺失。另一个原因是PCR使用了不同的靶基因。核糖体DNA(rDNA)簇的内转录间隔区(ITS)序列推荐用于真菌的DNA条形码分析。在临床样本中使用DNA测序鉴定真菌存在一定局限性,并且在某些情况下结果的解释可能存在问题。DNA序列在互联网上的公共数据库中进行搜索和比较,其中最著名的是GenBank。然而,专门的真菌学数据库提供了更可靠的真菌鉴定数据。

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