Separation of phenotypically indistinguishable Candida species,
C. orthopsilosis, C. metapsilosis and C. parapsilosis, by
capillary electromigration techniques

J 2011

Separation of phenotypically indistinguishable Candida species, C. orthopsilosis, C. metapsilosis and C. parapsilosis, by capillary electromigration techniques

HORKÁ, Marie; Filip RŮŽIČKA; Anna KUBESOVÁ; Eva NĚMCOVÁ; Karel ŠLAIS et al.

Základní údaje

Originální název

Separation of phenotypically indistinguishable Candida species, C. orthopsilosis, C. metapsilosis and C. parapsilosis, by capillary electromigration techniques

Autoři

HORKÁ, Marie; Filip RŮŽIČKA ; Anna KUBESOVÁ; Eva NĚMCOVÁ a Karel ŠLAIS

Vydání

Journal of Chromatography A, 2011, 0021-9673

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 4.531

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14110/11:00049493

Organizační jednotka

Lékařská fakulta

DOI

https://doi.org/10.1016/j.chroma.2011.04.057

UT WoS

000291911000009

Klíčová slova anglicky

Capillary electromigration techniques with UV detection; Capillary isoelectric focusing in narrow pH gradient; Candida orthopsilosis; Candida metapsilosis; Candida parapsilosis

Příznaky

Mezinárodní význam

Změněno: 30. 6. 2012 17:23, MUDr. Mgr. Karel Šlais, Ph.D.

Anotace

V originále

At the current state of laboratory diagnostics, methods for fast identification of phenotypically indistinguishable species are difficult or inaccurate. An example is represented by Candida parapsilosis, which is the second most common yeast species isolated from bloodstream infections. C. parapsilosis comprises a complex of three genetically distinct groups. Genotypes II and III have been designated as the separate species Candida orthopsilosis and Candida metapsilosis, phenotypically indistinguishable. The considerable genetic variability of these newly described yeasts species has caused difficulties in the development of molecular techniques for their precise identification. Similarly, the detection of biofilm formation, which is considered as an important yeast virulence factor, is accompanied by difficulties. In this study we optimize the first precise and reproducible method for the separation and possible identification of C. orthopsilosis, C. metapsilosis and C. parapsilosis as well as the detection of their ability to form biofilm. The method is based on capillary isoelectric focusing and capillary electrophoresis with UV detection. In capillary isoelectric focusing, very narrow pH gradients were established. With such gradients, differences in isoelectric points of biofilm-negative and biofilm-positive species calculated from the migration times of the selected pl markers were below 0.03 pl units. In the capillary zone electrophoresis narrow zones of the cells of Candida species were detected with sufficient resolution. The values of the isoelectric point and the migration velocities of the examined species were independent on the origin of the tested strains. Capillary isoelectric focusing was examined also for the separation and detection of the cultivated biofilm-negative C parapsilosis in the blood serum.

Návaznosti

IAAX00310701, projekt VaV

Název: Rychlá detekce a identifikace patogenních mikroorganizmů a virů pomocí elektromigračních technik a hmotnostní spektrometrie

Investor: Akademie věd ČR, Rychlá detekce a identifikace patogenních mikroorganismů a virů pomocí elektromigračních technik a hmotnostní spektrometrie

1M0528, projekt VaV

Název: Stomatologické výzkumné centrum

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Stomatologické výzkumné centrum

Citovat

HORKÁ, Marie; Filip RŮŽIČKA; Anna KUBESOVÁ; Eva NĚMCOVÁ a Karel ŠLAIS. Separation of phenotypically indistinguishable Candida species, C. orthopsilosis, C. metapsilosis and C. parapsilosis, by capillary electromigration techniques. Journal of Chromatography A. 2011, roč. 1218, č. 25, s. 3900-3907. ISSN 0021-9673. Dostupné z: https://doi.org/10.1016/j.chroma.2011.04.057.

@article{959071,
   author = {Horká, Marie and Růžička, Filip and Kubesová, Anna and Němcová, Eva and Šlais, Karel},
   article_number = {25},
   doi = {https://doi.org/10.1016/j.chroma.2011.04.057},
   keywords = {Capillary electromigration techniques with UV detection; Capillary isoelectric focusing in narrow pH gradient; Candida orthopsilosis; Candida metapsilosis; Candida parapsilosis},
   language = {eng},
   issn = {0021-9673},
   journal = {Journal of Chromatography A},
   title = {Separation of phenotypically indistinguishable Candida species, C. orthopsilosis, C. metapsilosis and C. parapsilosis, by capillary electromigration techniques},
   volume = {1218},
   year = {2011}
}

TY  - JOUR
ID  - 959071
AU  - Horká, Marie - Růžička, Filip - Kubesová, Anna - Němcová, Eva - Šlais, Karel
PY  - 2011
TI  - Separation of phenotypically indistinguishable Candida species, C. orthopsilosis, C. metapsilosis and C. parapsilosis, by capillary electromigration techniques
JF  - Journal of Chromatography A
VL  - 1218
IS  - 25
SP  - 3900-3907
EP  - 3900-3907
SN  - 00219673
KW  - Capillary electromigration techniques with UV detection
KW  - Capillary isoelectric focusing in narrow pH gradient
KW  - Candida orthopsilosis
KW  - Candida metapsilosis
KW  - Candida parapsilosis
N2  - At the current state of laboratory diagnostics, methods for fast identification of phenotypically indistinguishable species are difficult or inaccurate. An example is represented by Candida parapsilosis, which is the second most common yeast species isolated from bloodstream infections. C. parapsilosis comprises a complex of three genetically distinct groups. Genotypes II and III have been designated as the separate species Candida orthopsilosis and Candida metapsilosis, phenotypically indistinguishable. The considerable genetic variability of these newly described yeasts species has caused difficulties in the development of molecular techniques for their precise identification. Similarly, the detection of biofilm formation, which is considered as an important yeast virulence factor, is accompanied by difficulties. In this study we optimize the first precise and reproducible method for the separation and possible identification of C. orthopsilosis, C. metapsilosis and C. parapsilosis as well as the detection of their ability to form biofilm. The method is based on capillary isoelectric focusing and capillary electrophoresis with UV detection. In capillary isoelectric focusing, very narrow pH gradients were established. With such gradients, differences in isoelectric points of biofilm-negative and biofilm-positive species calculated from the migration times of the selected pl markers were below 0.03 pl units. In the capillary zone electrophoresis narrow zones of the cells of Candida species were detected with sufficient resolution. The values of the isoelectric point and the migration velocities of the examined species were independent on the origin of the tested strains. Capillary isoelectric focusing was examined also for the separation and detection of the cultivated biofilm-negative C parapsilosis in the blood serum.
ER  -

HORKÁ, Marie; Filip RŮŽIČKA; Anna KUBESOVÁ; Eva NĚMCOVÁ a Karel ŠLAIS. Separation of phenotypically indistinguishable Candida species, C. orthopsilosis, C. metapsilosis and C. parapsilosis, by capillary electromigration techniques. \textit{Journal of Chromatography A}. 2011, roč.~1218, č.~25, s.~3900-3907. ISSN~0021-9673. Dostupné z: https://doi.org/10.1016/j.chroma.2011.04.057.

Přehled o publikaci