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学科主题: 临床医学
题名:
Rapid detection of pathogenic fungi using loop-mediated isothermal amplification, exemplified by Fonsecaea agents of chromoblastomycosis
作者: Sun, Jiufeng1; Najafzadeh, M. J.2,3,4; Vicente, V.5; Xi, Liyan1; de Hoog, G. S.1,2,6
关键词: Chromoblastomycosis ; Fonsecaea ; LAMP ; Microbial detection
刊名: JOURNAL OF MICROBIOLOGICAL METHODS
发表日期: 2010
DOI: 10.1016/j.mimet.2009.10.002
卷: 80, 期:1, 页:19-24
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]: POLYMERASE-CHAIN-REACTION ; OLIGONUCLEOTIDE PRIMERS ; EXOPHIALA-SPINIFERA ; MYCOBACTERIUM-TUBERCULOSIS ; PHIALOPHORA-VERRUCOSA ; CAUSATIVE AGENT ; GENUS FONSECAEA ; DNA-SEQUENCES ; LAMP METHOD ; IDENTIFICATION
英文摘要:

Loop-mediated isothermal amplification (LAMP) was developed for rapid detection of pathogenic or allergenic fungal in the environment. Primers applied were derived from the rDNA Internal Transcribed Spacer and the 5.8S rRNA gene. The assay enabled amplification of target fungi at the level of genus or closely related species using pure cultures after I h reaction at 65 degrees C in a water bath. No cross-reactivity to related species was observed. The DNA detection limit was 0.2 fg. The method also proved to work well with fungi on non-sterile adhesive tape. Amplification products were detected by visual inspection using SYBR Green I as well as by electrophoresis on agarose gels. As a model organism we selected Fonsecaea, a fungal genus containing etiologic agents of chromoblastomycosis, a widely distributed tropical and subtropical skin disease in otherwise healthy patients and supposed to be acquired by environmental inoculation. It is suggested that LAMP can also be used for rapid clinical diagnosis, for environmental detection, and for retrospective studies in archived clinical samples. (C) 2009 Elsevier B.V. All rights reserved.

语种: 英语
所属项目编号: 200802026 ; 017626
项目资助者: International Program of Project 985 ; Ministry of Health of the People&prime ; s Republic of China ; EU
WOS记录号: WOS:000273846600003
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/65347
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.CBS KNAW Fungal Biodivers Ctr, Utrecht, Netherlands
2.Mashhad Univ Med Sci, Dept Parasitol & Mycol, Mashhad, Iran
3.Sun Yat Sen Univ, Affiliated Hosp 2, Dept Dermatol, Guangzhou 510120, Guangdong, Peoples R China
4.Univ Amsterdam, Inst Biodivers & Ecosyst Dynam, NL-1012 WX Amsterdam, Netherlands
5.Univ Fed Parana, Dept Basic Pathol, BR-80060000 Curitiba, Parana, Brazil
6.Peking Univ, Hlth Sci Ctr, Res Ctr Med Mycol, Beijing 100871, Peoples R China

Recommended Citation:
Sun, Jiufeng,Najafzadeh, M. J.,Vicente, V.,et al. Rapid detection of pathogenic fungi using loop-mediated isothermal amplification, exemplified by Fonsecaea agents of chromoblastomycosis[J]. JOURNAL OF MICROBIOLOGICAL METHODS,2010,80(1):19-24.
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