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学科主题: 临床医学
题名:
Detection and identification of opportunistic Exophiala species using the rolling circle amplification of ribosomal internal transcribed spacers
作者: Najafzadeh, M. J.1,2,3; Dolatabadi, S.3; Keisari, M. Saradeghi3,4; Naseri, A.5; Feng, P.3,6; de Hoog, G. S.3,7,8,9,10,11,12
关键词: Exophiala ; Black yeasts ; Identification ; Rolling circle amplification
刊名: JOURNAL OF MICROBIOLOGICAL METHODS
发表日期: 2013-09-01
DOI: 10.1016/j.mimet.2013.06.026
卷: 94, 期:3, 页:338-342
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Microbiology
研究领域[WOS]: Biochemistry & Molecular Biology ; Microbiology
关键词[WOS]: IN-VITRO ACTIVITIES ; RAPID IDENTIFICATION ; ANTIFUNGAL DRUGS ; HUMAN CHROMOBLASTOMYCOSIS ; FUNGAL PATHOGENS ; UNITED-STATES ; DERMATITIDIS ; FONSECAEA ; AGENT ; CYPHELLOPHORA
英文摘要:

Deep infections by melanized fungi deserve special attention because of a potentially fatal, cerebral or disseminated course of disease in otherwise healthy patients. Timely diagnostics are a major problem with these infections. Rolling circle amplification (RCA) is a sensitive, specific and reproducible isothermal DNA amplification technique for rapid molecular identification of microorganisms. RCA-based diagnostics are characterized by good reproducibility, with few amplification errors compared to PCR. The method is applied here to species of Exophiala known to cause systemic infections in humans. The ITS rDNA region of five Exophiala species (E. dermatitidis, E. oligosperma, E. spinifera, E. xenobiotica, and E. jeanselmei) was sequenced and aligned in view of designing specific padlock probes to be used for the detection of single nucleotide polymorphisms (SNPs) of the Exophiala species concerned. The assay proved to successfully amplify DNA of the target fungi at the level of species; while no cross-reactivity was observed. Amplification products were visualized on 1% agarose gels to verify the specificity of probe-template binding. Amounts of reagents were minimized to avoid the generation of false positive results. The sensitivity of RCA may help to improve early diagnostics of these difficult to diagnose infections. (C) 2013 Elsevier B.V. All rights reserved.

语种: 英语
所属项目编号: 920110
项目资助者: Mashhad University of Medical Sciences, Mashhad, Iran
WOS记录号: WOS:000324664500028
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/50365
Appears in Collections:北京大学第一临床医学院_皮肤性病科_期刊论文

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作者单位: 1.Univ Amsterdam, Inst Biodivers & Ecosyst Dynam, Amsterdam, Netherlands
2.Mashhad Univ Med Sci, Sch Med, Ghaem Hosp, Dept Parasitol & Mycol, Mashhad, Iran
3.Mashhad Univ Med Sci, Sch Med, Ghaem Hosp, Canc Mol Pathol Res Ctr, Mashhad, Iran
4.CBS KNAW Fungal Biodivers Ctr, Utrecht, Netherlands
5.Mashhad Univ Med Sci, Food & Drug Adm, Mashhad, Iran
6.Mashhad Univ Med Sci, Sch Med, Imam Reza Hosp, Dept Parasitol & Mycol, Mashhad, Iran
7.Sun Yat Sen Univ, Affiliated Hosp 3, Guangzhou 510275, Guangdong, Peoples R China
8.Sun Yat Sen Univ, Sun Yat Sen Mem Hosp, Guangzhou 510275, Guangdong, Peoples R China
9.Peking Univ, Hlth Sci Ctr, Res Ctr Med Mycol, Beijing 100871, Peoples R China
10.Second Mil Med Univ, Changzheng Hosp, Shanghai Inst Med Mycol, Shanghai, Peoples R China
11.Univ Fed Parana, Basic Pathol Dept, Curitiba, Parana, Brazil
12.King Abdulassiz Univ, Jeddah, Saudi Arabia

Recommended Citation:
Najafzadeh, M. J.,Dolatabadi, S.,Keisari, M. Saradeghi,et al. Detection and identification of opportunistic Exophiala species using the rolling circle amplification of ribosomal internal transcribed spacers[J]. JOURNAL OF MICROBIOLOGICAL METHODS,2013,94(3):338-342.
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