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珊瑚中与白瘟病相关的活性弧菌的定量检测

Quantitative Detection of Active Vibrios Associated with White Plague Disease in Corals.

作者信息

Chimetto Tonon Luciane A, Thompson Janelle R, Moreira Ana P B, Garcia Gizele D, Penn Kevin, Lim Rachelle, Berlinck Roberto G S, Thompson Cristiane C, Thompson Fabiano L

机构信息

Laboratory of Organic Chemistry of Biological Systems, Chemical Institute of São Carlos, University of São Paulo, São Carlos, Brazil.

Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States.

出版信息

Front Microbiol. 2017 Nov 17;8:2272. doi: 10.3389/fmicb.2017.02272. eCollection 2017.

Abstract

Over recent decades several coral diseases have been reported as a significant threat to coral reef ecosystems causing the decline of corals cover and diversity around the world. The development of techniques that improve the ability to detect and quantify microbial agents involved in coral disease will aid in the elucidation of disease cause, facilitating coral disease detection and diagnosis, identification and pathogen monitoring, pathogen sources, vectors, and reservoirs. The genus is known to harbor pathogenic strains to marine organisms. One of the best-characterized coral pathogens is , an aetilogic agent of White Plague Disease (WPD). We used coral tissue (healthy and diseased specimens) to develop a rapid reproducible detection system for vibrios based on RT-QPCR and SYBR chemistry. We were able to detect total vibrios in expressed RNA targeting the 16S rRNA gene at 5.23 × 10 copies/μg RNA and targeting the gene at 5.10 × 10 copies/μg RNA in coral tissue. Detection of in diseased and in healthy samples suggests that WPD in the Abrolhos Bank may be caused by a consortium of microorganism and not only a single pathogen. We developed a more practical and economic system compared with probe uses for the real-time detection and quantification of vibrios from coral tissues by using the 16S rRNA and pyrH gene. This qPCR assay is a reliable tool for the monitoring of coral pathogens, and can be useful to prevent, control, or reduce impacts in this ecosystem.

摘要

近几十年来,有报道称几种珊瑚疾病对珊瑚礁生态系统构成重大威胁,导致全球珊瑚覆盖率和多样性下降。开发能够提高检测和量化参与珊瑚疾病的微生物病原体能力的技术,将有助于阐明疾病成因,促进珊瑚疾病的检测与诊断、病原体鉴定与监测、病原体来源、传播媒介和宿主的研究。已知该属含有对海洋生物致病的菌株。最具特征的珊瑚病原体之一是,它是白瘟病(WPD)的病原体。我们使用珊瑚组织(健康和患病标本),基于RT-QPCR和SYBR化学方法开发了一种快速可重复的弧菌检测系统。我们能够在珊瑚组织中检测到靶向16S rRNA基因的表达RNA中的总弧菌,其含量为5.23×10拷贝/μg RNA,靶向基因的含量为5.10×10拷贝/μg RNA。在患病和健康样本中检测到表明,阿布洛霍斯浅滩的WPD可能是由微生物群落引起的,而不仅仅是单一病原体。与使用探针实时检测和量化珊瑚组织中的弧菌相比,我们开发了一种更实用、更经济的系统,该系统使用16S rRNA和pyrH基因。这种qPCR检测方法是监测珊瑚病原体的可靠工具,可用于预防、控制或减少对该生态系统的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/39d7/5698304/63f4f38d2ce0/fmicb-08-02272-g0001.jpg

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