Electromigration techniques – a fast and economical tool for differentiation of similar strains of microorganisms

Abstract

The detection and identification of pathogens currently relies upon a very diverse range of techniques and skills, from traditional cultivation and taxonomic procedures to modern rapid and sensitive diagnostic methods. Real-time PCR is now exploited as a front line diagnostic screening tool in human, animal and plant health as well as bio-security. Nevertheless, new techniques for pathogen identification, particularly of unknown samples, are needed. In this study we propose the combination of electrophoresis-based procedures for the fast differentiation of microorganisms. The method feasibility is proved on the model of seven similar strains of Pseudomonas syringae pathovars from 37 sources, identified by gas chromatography of fatty acid methyl esters. The results from the routine laboratory were compared with results of the combination of the developed capillary and gel electrophoresis as well as mass spectrometry. According to our experiments appropriate combination of electromigration techniques appears to be useful for the fast and economical differentiation of unknown microorganisms.

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

“…Overall, although the application of CE based on nucleic acid detection has its limitations, the many advantages over the traditional culture-based, phenotype and biochemical test identification still attract many researchers to this approach [33,152]. The development of this technique has been and the future developments are likely to be as follows: 1.…”

Direct and Indirect Applications of Sub- and Supercritical Water in Food-Related Analysis

“…The reliability of identification can be improved by using the matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF MS) (Demirev and Fenselau 2008;Carbonnelle et al 2011). This option adds a powerful off-line identification dimension to the CIEF separation and focusing as the individual microbial species usually display their respective individual characteristic mass spectrometric fingerprints (Horká et al 2010;Š alplachta et al 2013).…”

Recent developments in capillary and chip electrophoresis of bioparticles: Viruses, organelles, and cells

“…In the most recent paper [41], Horká et al proposed a combination of CZE and CIEF for the differentiation of unknown microorganisms, considering costs and speed of analysis. For this purpose, seven similar strains of P. syringae pathovars from 37 different sources and a strain of P. corrugata were analyzed by these electromigration techniques, as well as by gel IEF, GC of fatty acid methyl esters, and MALDI-TOF MS. For CZE and CIEF, PEG as dynamic modifier was employed; CIEF was performed in the pH range of 2.0-4.7.…”