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@article{1623016,
author = {Horka, Marie and Salplachta, Jiri and Růžička, Filip and Slais, Karel},
article_location = {WASHINGTON},
article_number = {8},
doi = {http://dx.doi.org/10.1021/acsinfecdis.9b00045},
keywords = {uropathogens; high level of albumin; red nonionogenic tenside; preconcentration and preseparation; isoelectric focusing; matrix-assisted laser desorption/ionization time-of-flight mass spectrometry},
language = {eng},
issn = {2373-8227},
journal = {ACS INFECTIOUS DISEASES},
title = {Utilization of Red Nonionogenic Tenside Labeling, Isoelectric Focusing, and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry in the Identification of Uropathogens in the Presence of a High Level of Albumin},
url = {http://dx.doi.org/10.1021/acsinfecdis.9b00045},
volume = {5},
year = {2019}
}
TY - JOUR
ID - 1623016
AU - Horka, Marie - Salplachta, Jiri - Růžička, Filip - Slais, Karel
PY - 2019
TI - Utilization of Red Nonionogenic Tenside Labeling, Isoelectric Focusing, and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry in the Identification of Uropathogens in the Presence of a High Level of Albumin
JF - ACS INFECTIOUS DISEASES
VL - 5
IS - 8
SP - 1348-1356
EP - 1348-1356
PB - AMER CHEMICAL SOC
SN - 23738227
KW - uropathogens
KW - high level of albumin
KW - red nonionogenic tenside
KW - preconcentration and preseparation
KW - isoelectric focusing
KW - matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
UR - http://dx.doi.org/10.1021/acsinfecdis.9b00045
L2 - http://dx.doi.org/10.1021/acsinfecdis.9b00045
N2 - Cellulose-based preparative isoelectric focusing was used for preseparation and concentration of uropathogens Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Staphylococcus epidermidis, Candida albicans, and Candida parapsilosis in a urine sample containing a high concentration of human serum albumin. For the visibility of the colorless microbial zones in the separation medium, the microbial cells were labeled with red nonionogenic tenside (1-[[4-(phenylazo)-phenyl]azo]-2-hydroxy-3-naphthoic acid polyethylene glycol ester, PAPAN). A very short incubation time, about 2 min, was sufficient for the adsorption of 0.001% (w/v) PAPAN onto the cell surface at the optimized conditions. As low as 10(3) cells of E. coli (pI 4.6) resuspended in 100 mu L of urine sample and spiked with 0.1 mg mL(-1) of human serum albumin (pI 4.8) were successfully preseparated and concentrated using this method. Because the pI values of the labeled microorganisms remained unchanged, the focused red zones of microbial cells were collected from the separation media and further analyzed by either capillary isoelectric focusing or matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The viability of the cells extracted from the collected zones was also confirmed. The proposed method provides reliable, relatively fast, and cost-effective identification of uropathogens in urine specimens with a high level of albumin.
ER -
HORKA, Marie, Jiri SALPLACHTA, Filip RŮŽIČKA and Karel SLAIS. Utilization of Red Nonionogenic Tenside Labeling, Isoelectric Focusing, and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry in the Identification of Uropathogens in the Presence of a High Level of Albumin. \textit{ACS INFECTIOUS DISEASES}. WASHINGTON: AMER CHEMICAL SOC, 2019, vol.~5, No~8, p.~1348-1356. ISSN~2373-8227. Available from: https://dx.doi.org/10.1021/acsinfecdis.9b00045.
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