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 identification of fungal pathogens by rolling circle amplification using Fonsecaea as a model
Najafzadeh, M. J.1,2,3; Sun, J.2,4; Vicente, V. A.2,5; de Hoog, G. S.1,2,6
关键词Fonsecaea chromoblastomycosis RCA rapid identification
刊名MYCOSES
2011-09-01
DOI10.1111/j.1439-0507.2010.01995.x
54期:5页:E577-E582
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Dermatology ; Mycology
资助者Ministry of Health and Medical Education of Iran ; Mashhad University of Medical Sciences, Mashhad, Iran ; Coordenacao de Pessoal de Nivel Superior (CAPES) ; Ministry of Health and Medical Education of Iran ; Mashhad University of Medical Sciences, Mashhad, Iran ; Coordenacao de Pessoal de Nivel Superior (CAPES)
研究领域[WOS]Dermatology ; Mycology
关键词[WOS]PADLOCK PROBES ; PRACTICAL METHOD ; CHROMOBLASTOMYCOSIS ; OLIGONUCLEOTIDES ; POLYMORPHISMS ; REPLICATION
英文摘要

We aimed to describe a rapid and sensitive assay for identification of pathogenic fungi without sequencing. The method of rolling circle amplification (RCA) is presented with species of Fonsecaea, agents of human chromoblastomycosis, as a model. The internal transcribed spacer (ITS) rDNA region of 103 Fonsecaea strains was sequenced and aligned in view of designing three specific padlock probes to be used for the detection of single nucleotide polymorphisms in three Fonsecaea species. The 38 strains included for testing the specificity of RCA comprised 17 isolates of Fonsecaea pedrosoi, 1.3 of Fonsecaea monophora and eight of Fonsecaea nubica. The assay successfully amplified DNA of the target fungi at the level of species, while no cross reactivity was observed. The amplification product was visualised on a 1% agarose gel to verify the specificity of probe-template binding. Amounts of reagents were minimised to avoid the generation of false-positive results. The simplicity, sensitivity, robustness and low costs provide RCA a distinct position among isothermal techniques for DNA diagnostics as a very practical identification method.

语种英语
资助者Ministry of Health and Medical Education of Iran ; Mashhad University of Medical Sciences, Mashhad, Iran ; Coordenacao de Pessoal de Nivel Superior (CAPES) ; Ministry of Health and Medical Education of Iran ; Mashhad University of Medical Sciences, Mashhad, Iran ; Coordenacao de Pessoal de Nivel Superior (CAPES)
WOS记录号WOS:000294878900135
Citation statistics
Cited Times:20[WOS]   [WOS Record]     [Related Records in WOS]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/66488
Collection北京大学第一临床医学院_皮肤性病科
作者单位1.Univ Amsterdam, Inst Biodivers & Ecosyst Dynam, NL-1012 WX Amsterdam, Netherlands
2.CBS KNAW Fungal Biodivers Ctr, Utrecht, Netherlands
3.Mashhad Univ Med Sci, Fac Med, Dept Parasitol & Mycol, Mashhad, Iran
4.Sun Yat Sen Univ, Affiliated Hosp 2, Dept Dermatol, Guangzhou 510275, Guangdong, Peoples R China
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
GB/T 7714
Najafzadeh, M. J.,Sun, J.,Vicente, V. A.,et al. Rapid identification of fungal pathogens by rolling circle amplification using Fonsecaea as a model[J]. MYCOSES,2011,54(5):E577-E582.
APA Najafzadeh, M. J.,Sun, J.,Vicente, V. A.,&de Hoog, G. S..(2011).Rapid identification of fungal pathogens by rolling circle amplification using Fonsecaea as a model.MYCOSES,54(5),E577-E582.
MLA Najafzadeh, M. J.,et al."Rapid identification of fungal pathogens by rolling circle amplification using Fonsecaea as a model".MYCOSES 54.5(2011):E577-E582.
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