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学科主题临床医学
Rapid identification and characterization of Penicillium marneffei using multiplex ligation-dependent probe amplification (MLPA) in paraffin-embedded tissue samples
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
2011-04-01
DOI10.1016/j.mimet.2011.01.022
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
资助者National Natural Science Foundation of China ; Guangdong Natural Science Fund ; National Natural Science Foundation of China ; Guangdong Natural Science Fund
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
资助者National Natural Science Foundation of China ; Guangdong Natural Science Fund ; National Natural Science Foundation of China ; Guangdong Natural Science Fund
WOS记录号WOS:000289389600006
引用统计
被引频次:8[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/400002259/59017
专题北京大学第一临床医学院_皮肤性病科
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
推荐引用方式
GB/T 7714
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.
APA Zhang, Jun-Min.,Sun, Jiu-Feng.,Feng, Pei-Ying.,Li, Xi-Qing.,Lu, Chang-Ming.,...&de Hoog, G. S..(2011).Rapid identification and characterization of Penicillium marneffei using multiplex ligation-dependent probe amplification (MLPA) in paraffin-embedded tissue samples.JOURNAL OF MICROBIOLOGICAL METHODS,85(1),33-39.
MLA Zhang, Jun-Min,et al."Rapid identification and characterization of Penicillium marneffei using multiplex ligation-dependent probe amplification (MLPA) in paraffin-embedded tissue samples".JOURNAL OF MICROBIOLOGICAL METHODS 85.1(2011):33-39.
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