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.

Basic information

Original name

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

Authors

HORKÁ, Marie (203 Czech Republic, guarantor), Filip RŮŽIČKA (203 Czech Republic, belonging to the institution), Anna KUBESOVÁ (203 Czech Republic, belonging to the institution), Eva NĚMCOVÁ (203 Czech Republic, belonging to the institution) and Karel ŠLAIS (203 Czech Republic, belonging to the institution)

Edition

Journal of Chromatography A, 2011, 0021-9673

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 4.531

RIV identification code

RIV/00216224:14110/11:00049493

Organization unit

Faculty of Medicine

DOI

UT WoS

000291911000009

Keywords in English

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

Tags

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

Abstract

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.

Links

IAAX00310701, research and development project
Name: Rychlá detekce a identifikace patogenních mikroorganizmů a virů pomocí elektromigračních technik a hmotnostní spektrometrie
Investor: Academy of Sciences of the Czech Republic
1M0528, research and development project
Name: Stomatologické výzkumné centrum
Investor: Ministry of Education, Youth and Sports of the CR, Dental research centre