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@article{1674768,
author = {Horka, Marie and Salplachta, Jiri and Karasek, Pavel and Růžička, Filip and Roth, Michal},
article_location = {WASHINGTON},
article_number = {3},
doi = {http://dx.doi.org/10.1021/acsinfecdis.9b00200},
keywords = {capillary electrophoresis; matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; roughened capillary; cell-surface adhesion; Staphylococcus aureus; Pseudomonas aeruginosa},
language = {eng},
issn = {2373-8227},
journal = {ACS INFECTIOUS DISEASES},
title = {Online Concentration of Bacteria from Tens of Microliter Sample Volumes in Roughened Fused Silica Capillary with Subsequent Analysis by Capillary Electrophoresis and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry},
url = {https://pubs.acs.org/doi/10.1021/acsinfecdis.9b00200#},
volume = {6},
year = {2020}
}
TY - JOUR
ID - 1674768
AU - Horka, Marie - Salplachta, Jiri - Karasek, Pavel - Růžička, Filip - Roth, Michal
PY - 2020
TI - Online Concentration of Bacteria from Tens of Microliter Sample Volumes in Roughened Fused Silica Capillary with Subsequent Analysis by Capillary Electrophoresis and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry
JF - ACS INFECTIOUS DISEASES
VL - 6
IS - 3
SP - 355-365
EP - 355-365
PB - AMER CHEMICAL SOC
SN - 23738227
KW - capillary electrophoresis
KW - matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
KW - roughened capillary
KW - cell-surface adhesion
KW - Staphylococcus aureus
KW - Pseudomonas aeruginosa
UR - https://pubs.acs.org/doi/10.1021/acsinfecdis.9b00200#
L2 - https://pubs.acs.org/doi/10.1021/acsinfecdis.9b00200#
N2 - This study presents a timely, reliable, and sensitive method for identification of pathogenic bacteria in clinical samples based on a combination of capillary electrophoresis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In this respect, a part of a single-piece fused silica capillary was etched with supercritical water with the aim of using it for static or dynamic cell-surface adhesion from tens of microliter sample volumes. The conditions for this procedure were optimized. Adhered cells of Staphylococcus aureus (methicillinsusceptible or methicillin-resistant) and of Pseudomonas aeruginosa were desorbed and preconcentrated from the rough part of the capillary surface using transient isotachophoretic stacking from a high conductivity model matrix. The charged cells were swep and separated again in micellar electrokinetic chromatography using a nonionogenic surfactant. Static adhesion of the cells onto the roughened part of the capillary is certainly volumetric limited. Dynamic adhesion allows the concentration of bacteria from 100 mu L volumes of physiological saline solution, bovine serum, or human blood with the limits of detection at 1.8 x 10(2), 1.7 x 10(3), and 1.0 x 10(3) cells mL(-1), respectively. The limits of detection were the same for all three examined bacterial strains. The recovery of the method was about 83% and it was independent of the sample matrix. A combination of capillary electrophoresis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry required at least 4 x 103 cells mL-1 to obtain reliable results. The calibration plots were linear (R-2 = 0.99) and the relative standard deviations of the peak area were at most 2.2%. The adhered bacteria, either individual or in a mixture, were online analyzed by micellar electrokinetic chromatography and then collected from the capillary and off-line analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry without interfering matrix components.
ER -
HORKA, Marie, Jiri SALPLACHTA, Pavel KARASEK, Filip RŮŽIČKA a Michal ROTH. Online Concentration of Bacteria from Tens of Microliter Sample Volumes in Roughened Fused Silica Capillary with Subsequent Analysis by Capillary Electrophoresis and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry. \textit{ACS INFECTIOUS DISEASES}. WASHINGTON: AMER CHEMICAL SOC, roč.~6, č.~3, s.~355-365. ISSN~2373-8227. doi:10.1021/acsinfecdis.9b00200. 2020.
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