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IR@PKUHSC  > 北京大学第一临床医学院  > 皮肤性病科  > 期刊论文
学科主题: 临床医学
题名:
Rapid identification and characterization of Penicillium marneffei using multiplex ligation-dependent probe amplification (MLPA) in paraffin-embedded tissue samples
作者: Zhang, Jun-Min1; Sun, Jiu-Feng1,2; Feng, Pei-Ying1,3; Li, Xi-Qing1; Lu, Chang-Ming1; Lu, Sha1; Cai, Wen-Ying1; Xi, Li-Yan1; de Hoog, G. S.1,4,5
关键词: Penicillium marneffei ; Penicilliosis marneffei ; Fungal infection ; Multiplex ligation-dependent probe amplification ; MLPA
刊名: JOURNAL OF MICROBIOLOGICAL METHODS
发表日期: 2011-04-01
DOI: 10.1016/j.mimet.2011.01.022
卷: 85, 期:1, 页:33-39
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 ; BOVINE SERUM-ALBUMIN ; RELATIVE QUANTIFICATION ; QUANTITATIVE METHOD ; PADLOCK PROBES ; NESTED PCR ; GENE ; SEQUENCES ; INFECTION ; ASSAY
英文摘要:

Penicillium marneffei infection is a deadly disease and early diagnosis leads to prompt and appropriate antifungal therapy. To develop a sensitive method to diagnose P. morneffei infection, a multiplex ligation-dependent probe amplification (MLPA) assay was adapted. This method can rapidly and specifically detect P. marneffei DNA in cultured cells and paraffin-embedded tissue samples. Three pairs of probes were designed for amplifying the internally (intergenic) transcribed spacer (ITS) region of P. marneffei rRNA using a systematic phylogenetic analysis. These three probe sets produced three amplicons of 198, 166, and 152 bp, respectively, specific for P. marneffei. In contrast, there was only one 198 bp amplicon produced for Talaromyces stipitatus, and one 152 bp amplicon for P. funiculosum, T. intermedius and T. derxii. The probes did not amplify any other reference strains. An array of 40 P. marneffei strains isolated from human patients, bamboo rat, and the local environment was tested by using MLPA, and all were positively identified. Most importantly, P. marneffei in paraffin-embedded tissue specimens from infected human patients was positively amplified by MLPA. The sensitivity and specificity of the MLPA assay could be a useful tool for prompt diagnosis, pathogen characterization, and epidemiological studies of fungal infections. (C) 2011 Elsevier B.V. All rights reserved.

语种: 英语
所属项目编号: 30770121/2007 ; 8151008901000073
项目资助者: National Natural Science Foundation of China ; Guangdong Natural Science Fund
WOS记录号: WOS:000289389600006
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/59017
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.Sun Yat Sen Univ, Affiliated Hosp 2, Dept Dermatol, Guangzhou 510120, Guangdong, Peoples R China
2.Sun Yat Sen Univ, Zhongshan Sch Med, Dept Parasitol, Guangzhou 510120, Guangdong, Peoples R China
3.Sun Yat Sen Univ, Affiliated Hosp 3, Dept Dermatol, Guangzhou 510120, Guangdong, Peoples R China
4.Univ Amsterdam, Inst Biodivers & Ecosyst Dynam, NL-1012 WX Amsterdam, Netherlands
5.Peking Univ, Hlth Sci Ctr, Res Ctr Med Mycol, Beijing 100871, Peoples R China

Recommended Citation:
Zhang, Jun-Min,Sun, Jiu-Feng,Feng, Pei-Ying,et al. Rapid identification and characterization of Penicillium marneffei using multiplex ligation-dependent probe amplification (MLPA) in paraffin-embedded tissue samples[J]. JOURNAL OF MICROBIOLOGICAL METHODS,2011,85(1):33-39.
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