RESUMO
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.
Assuntos
Bactérias/classificação , Bactérias/isolamento & purificação , Polietileno/química , Polietilenoimina/química , Microbiologia do Solo , Análise Espectral Raman/métodos , Bactérias/química , Aderência Bacteriana , Interações Hidrofóbicas e Hidrofílicas , Concentração Osmolar , Eletricidade EstáticaRESUMO
An easy and inexpensive detection method for DNA hybridization assays combining magnetic beads and enzymatically generated silver nanoparticles is introduced. The main advantage of this approach is the possibility to distinguish between positive and negative test results with the naked eye. In the case of complementary DNA sequences the sample will turn black within a few minutes, allowing readout without any hardware. In order to illustrate the applicability of the assay genomic DNA isolated from E. coli contaminated Ringer's solution was used for testing the sensitivity as well as specificity.