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通过酶联免疫吸附测定法(ELISA)以及超高效液相色谱-质谱联用(UHPLC-MS)检测细菌群体感应信号分子N-酰基高丝氨酸内酯(HSL)和N-酰基高丝氨酸(HS) 。

Detection of the Bacterial Quorum-Sensing Signaling Molecules N-Acyl-Homoserine Lactones (HSL) and N-Acyl-Homoserine (HS) with an Enzyme-Linked Immunosorbent Assay (ELISA) and via Ultrahigh-Performance Liquid Chromatography Coupled to Mass Spectrometry (UHPLC-MS).

作者信息

Rothballer Michael, Uhl Jenny, Kunze Josie, Schmitt-Kopplin Philippe, Hartmann Anton

机构信息

Research Unit Microbe-Plant Interactions, Department Environmental Sciences, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg, Germany.

Research Unit Analytical BioGeoChemistry, Department Environmental Sciences, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg, Germany.

出版信息

Methods Mol Biol. 2018;1673:61-72. doi: 10.1007/978-1-4939-7309-5_5.

Abstract

Quick and reliable quantitative methods requiring low amounts of sample volume are needed for the detection of N-acyl-homoserine lactones (HSL) and their degradation products N-acyl-homoserines (HS) in order to elucidate the occurrence and dynamics of these prevalent quorum-sensing molecules of Gram-negative bacteria in natural samples and laboratory model experiments. A combination of ELISA and UHPLC-MS is presented here which has proven to meet these requirements. Both methods can not only precisely detect and quantify HSLs but also their degradation products HS and thereby enable studying signaling dynamics in quorum sensing, which have been identified to play an essential role in bacterial communication.

摘要

为了阐明革兰氏阴性菌这些普遍存在的群体感应分子在天然样品和实验室模型实验中的出现情况和动态变化,需要快速可靠的定量方法,且所需样品体积少,用于检测N-酰基高丝氨酸内酯(HSL)及其降解产物N-酰基高丝氨酸(HS)。本文介绍了一种酶联免疫吸附测定(ELISA)和超高效液相色谱-质谱联用(UHPLC-MS)的组合方法,该方法已被证明能满足这些要求。这两种方法不仅能精确检测和定量HSL及其降解产物HS,还能研究群体感应中的信号动态变化,群体感应中的信号动态变化已被确定在细菌通讯中起着至关重要的作用。

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