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Legiolert™ 用于检测嗜肺军团菌的评估及其与平板划线培养法和定量聚合酶链反应法的比较。

Evaluation of Legiolert™ for the Detection of Legionella pneumophila and Comparison with Spread-Plate Culture and qPCR Methods.

作者信息

Monteiro Silvia N, Robalo Adriana M, Santos Ricardo J

机构信息

Laboratório de Analises, Universidade Lisboa, Instituto Superior Técnico, Lisbon, Portugal.

出版信息

Curr Microbiol. 2021 May;78(5):1792-1797. doi: 10.1007/s00284-021-02436-6. Epub 2021 Mar 23.

Abstract

Legionella pneumophila, the organism responsible for Legionnaires' disease, a potentially lethal pneumonia, is an opportunistic bacterium spread via inhalation of contaminated, aerosolized water. The detection and control of L. pneumophila is crucial to reduce the risk it poses to human health. L. pneumophila is generally detected and quantified by the plating method, ISO 11731:2017 and by qPCR. ISO 11731 is based on the filtration of the water sample through a membrane, which is placed on selective agar medium, and after colony growth, presumptive Legionella are then confirmed by subculturing, serology, or PCR. Quantitative Polymerase Chain Reaction (qPCR) is based on the amplification of a DNA sequence specific to L. pneumophila, usually within the mip gene. The objective of this study was to compare these methods to a new, liquid culture method based on the Most Probable Number (MPN) technique, Legiolert™/Quanti-Tray® with data obtained with ISO 11731 and a viability quantitative qPCR (v-qPCR), for quantification of L. pneumophila in potable and non-potable waters. Data showed that the Legiolert method revealed concentrations of L. pneumophila greater than ISO 11731 and generally similar results to those of v-qPCR. The Legiolert method was highly specific and easy to use, representing a significant advancement in the quantification of L. pneumophila from potable and non-potable waters.

摘要

嗜肺军团菌是导致军团病(一种潜在致命性肺炎)的病原体,它是一种通过吸入受污染的雾化水传播的机会致病菌。检测和控制嗜肺军团菌对于降低其对人类健康构成的风险至关重要。嗜肺军团菌通常通过平板计数法、ISO 11731:2017和定量聚合酶链反应(qPCR)进行检测和定量。ISO 11731基于将水样通过滤膜过滤,将滤膜置于选择性琼脂培养基上,菌落生长后,通过传代培养、血清学或PCR对疑似军团菌进行确认。定量聚合酶链反应(qPCR)基于对嗜肺军团菌特异性DNA序列的扩增,通常在mip基因内。本研究的目的是将这些方法与一种基于最大可能数(MPN)技术的新型液体培养方法Legiolert™/Quanti-Tray®进行比较,该方法与通过ISO 11731和活菌定量qPCR(v-qPCR)获得的数据一起,用于定量饮用水和非饮用水中的嗜肺军团菌。数据表明,Legiolert方法检测出的嗜肺军团菌浓度高于ISO 11731,且结果通常与v-qPCR相似。Legiolert方法具有高度特异性且易于使用,代表了饮用水和非饮用水中嗜肺军团菌定量检测方面的重大进展。

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