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一种开创性的基于颗粒的策略,用于从与拉曼光谱兼容的多部分土壤样本中分离活细菌。

Pioneering particle-based strategy for isolating viable bacteria from multipart soil samples compatible with Raman spectroscopy.

作者信息

Schwarz Martha, Kloß Sandra, Stöckel Stephan, Pollok Sibyll, Holländer Andreas, Cialla-May Dana, Weber Karina, Popp Jürgen

机构信息

Leibniz-Institute of Photonic Technology Jena, Albert-Einstein-Str. 9, 07745, Jena, Germany.

InfectoGnostics Forschungscampus Jena, Philosophenweg 7, 07743, Jena, Germany.

出版信息

Anal Bioanal Chem. 2017 Jun;409(15):3779-3788. doi: 10.1007/s00216-017-0320-z. Epub 2017 Mar 31.

DOI:10.1007/s00216-017-0320-z
PMID:28364142
Abstract

The study of edaphic bacteria is of great interest, particularly for evaluating soil remediation and recultivation methods. Therefore, a fast and simple strategy to isolate various bacteria from complex soil samples using poly(ethyleneimine) (PEI)-modified polyethylene particles is introduced. The research focuses on the binding behavior under different conditions, such as the composition, pH value, and ionic strength, of the binding buffer, and is supported by the characterization of the surface properties of particles and bacteria. The results demonstrate that electrostatic forces and hydrophobicity are responsible for the adhesion of target bacteria to the particles. Distinct advantages of the particle-based isolation strategy include simple handling, enrichment efficiency, and the preservation of viable bacteria. The presented isolation method allows a subsequent identification of the bacteria using Raman microspectroscopy in combination with chemometrical methods. This is demonstrated with a dataset of five different bacteria (Escherichia coli, Bacillus subtilis, Pseudomonas fluorescens, Streptomyces tendae, and Streptomyces acidiscabies) which were isolated from spiked soil samples. In total 92% of the Raman spectra could be identified correctly.

摘要

土壤细菌的研究具有重大意义,尤其是在评估土壤修复和复垦方法方面。因此,本文介绍了一种利用聚乙烯亚胺(PEI)改性聚乙烯颗粒从复杂土壤样品中分离各种细菌的快速简便策略。该研究聚焦于结合缓冲液在不同条件下(如组成、pH值和离子强度)的结合行为,并通过颗粒和细菌表面性质的表征得到支持。结果表明,静电力和疏水性是目标细菌与颗粒粘附的原因。基于颗粒的分离策略具有操作简单、富集效率高和能保存活细菌等明显优势。所提出的分离方法允许随后结合化学计量学方法使用拉曼光谱对细菌进行鉴定。从加标土壤样品中分离出的五种不同细菌(大肠杆菌、枯草芽孢杆菌、荧光假单胞菌、绛红链霉菌和酸腐链霉菌)的数据集证明了这一点。总共92%的拉曼光谱能够被正确识别。

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