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一种用于快速检测实验样本和临床样本中阿萨希毛孢子菌的环介导等温扩增检测方法的开发。

Development of a loop-mediated isothermal amplification assay for rapid detection of Trichosporon asahii in experimental and clinical samples.

作者信息

Zhou Jianfeng, Liao Yong, Li Haitao, Lu Xuelian, Han Xiufeng, Tian Yanli, Chen Shanshan, Yang Rongya

机构信息

Department of Dermatology, General Hospital of Beijing Military Command of PLA, No.5, Nanmencang, Dongcheng District, Beijing 100010, China ; The Clinical Medical College in the Beijing Military Region of Second Military Medical University of PLA, Beijing 100700, China.

Department of Dermatology, General Hospital of Beijing Military Command of PLA, No.5, Nanmencang, Dongcheng District, Beijing 100010, China.

出版信息

Biomed Res Int. 2015;2015:732573. doi: 10.1155/2015/732573. Epub 2015 Jan 27.

Abstract

Invasive trichosporonosis is a deep mycosis found mainly in immunocompromised hosts, and the major pathogen is Trichosporon asahii. We detected the species-specific intergenic spacers (IGS) of rRNA gene of T. asahii using a loop-mediated isothermal amplification (LAMP) assay in 15 isolates with 3 different visualization methods, including SYBR green detection, gel electrophoresis, and turbidimetric methods. The LAMP assay displayed superior rapidity to other traditional methods in the detection time; that is, only 1 h was needed for detection and identification of the pathogen DNA. Furthermore, the detection limit of the LAMP assay was more sensitive than the PCR assay. We also successfully detect the presence of T. asahii in samples from experimentally infected mice and samples from patients with invasive trichosporonosis caused by T. asahii, suggesting that this method may become useful in clinical applications in the near future.

摘要

侵袭性毛孢子菌病是一种主要在免疫功能低下宿主中发现的深部真菌病,主要病原体是阿萨希毛孢子菌。我们使用环介导等温扩增(LAMP)检测法,通过3种不同的可视化方法,即SYBR绿检测、凝胶电泳和比浊法,检测了15株阿萨希毛孢子菌rRNA基因的种特异性基因间隔区(IGS)。LAMP检测法在检测时间上比其他传统方法具有更高的快速性;也就是说,检测和鉴定病原体DNA仅需1小时。此外,LAMP检测法的检测限比聚合酶链反应(PCR)检测法更灵敏。我们还成功检测到了实验感染小鼠样本以及由阿萨希毛孢子菌引起的侵袭性毛孢子菌病患者样本中阿萨希毛孢子菌的存在,这表明该方法在不久的将来可能在临床应用中发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7457/4322298/b3adbd8174ac/BMRI2015-732573.001.jpg

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