IR@PKUHSC  > 北京大学第一临床医学院  > 皮肤性病科
学科主题临床医学
Rapid detection of pathogenic fungi using loop-mediated isothermal amplification, exemplified by Fonsecaea agents of chromoblastomycosis
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
2010
DOI10.1016/j.mimet.2009.10.002
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
WOS记录号WOS:000273846600003
项目编号200802026 ; 017626
资助机构International Program of Project 985 ; Ministry of Health of the People&prime ; s Republic of China ; EU
引用统计
被引频次:35[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/65347
专题北京大学第一临床医学院_皮肤性病科
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
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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.
APA Sun, Jiufeng,Najafzadeh, M. J.,Vicente, V.,Xi, Liyan,&de Hoog, G. S..(2010).Rapid detection of pathogenic fungi using loop-mediated isothermal amplification, exemplified by Fonsecaea agents of chromoblastomycosis.JOURNAL OF MICROBIOLOGICAL METHODS,80(1),19-24.
MLA Sun, Jiufeng,et al."Rapid detection of pathogenic fungi using loop-mediated isothermal amplification, exemplified by Fonsecaea agents of chromoblastomycosis".JOURNAL OF MICROBIOLOGICAL METHODS 80.1(2010):19-24.
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