HORKÁ, Marie, Filip RŮŽIČKA, Anna KUBEŠOVÁ and Karel ŠLAIS. Dynamic labeling of diagnostically significant microbial cells in cerebrospinal fluid by red chromophoric non-ionogenic surfactant for capillary electrophoresis separations. Analytica Chimica Acta. Amsterdam: Elsevier Science publishers, 2012, vol. 728, No 1, p. 86-92. ISSN 0003-2670. doi:10.1016/j.aca.2012.03.045.
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Basic information
Original name Dynamic labeling of diagnostically significant microbial cells in cerebrospinal fluid by red chromophoric non-ionogenic surfactant for capillary electrophoresis separations
Authors HORKÁ, Marie (203 Czech Republic, guarantor), Filip RŮŽIČKA (203 Czech Republic, belonging to the institution), Anna KUBEŠOVÁ (203 Czech Republic) and Karel ŠLAIS (203 Czech Republic).
Edition Analytica Chimica Acta, Amsterdam, Elsevier Science publishers, 2012, 0003-2670.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10600 1.6 Biological sciences
Country of publisher Netherlands
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 4.387
RIV identification code RIV/00216224:14110/12:00057148
Organization unit Faculty of Medicine
Doi http://dx.doi.org/10.1016/j.aca.2012.03.045
UT WoS 000304496000012
Keywords in English CZE and CIEF; Preconcentration; Microorganisms; Cerebrospinal fluid; Colored non-ionogenic surfactant; Dynamic modification
Tags International impact, Reviewed
Changed by Changed by: Ing. Mgr. Věra Pospíšilíková, učo 9005. Changed: 10. 2. 2013 14:47.
Abstract
During bacterial infections of the central nervous system the number of microorganisms in the cerebrospinal fluid is often ranging from few up to hundreds of cells per milliliter. The electrophoretic techniques with the UV-detection reach a detection limit for whole cells of approximately 10(7) cells per milliliter. The coupling of the filtration cartridge with capillary isoelectric focusing can improve the detection limit by four orders of magnitude. In order to improve the detection limit the red non-ionogenic surfactant 1-[[4-(phenylazo)phenyl]azo]-2-hydroxy-3-naphthoic acid polyethylene glycol ester, PAPAN 1000, has been prepared and used for the dynamic labeling of analytes before filtration of the sample with a concentration modulation in the analysis of proteins or microorganisms. Values of isoelectric points of labeled analytes have been calculated using pI markers detectable at 515 nm and have been found comparable with pi of the native compounds. Minimum detectable amounts of proteins and microorganisms were lower than nanograms and a hundred labeled cells, respectively. The introduced method, coupling of the filtration cerebrospinal fluid spiked with microorganisms and labeled by PAPAN, facilitates their rapid CIEF separation in the pH gradient pH range of 2-5 at their clinically important level 10(1) to 10(2) cells per milliliter.
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IAAX00310701, research and development projectName: Rychlá detekce a identifikace patogenních mikroorganizmů a virů pomocí elektromigračních technik a hmotnostní spektrometrie
Investor: Academy of Sciences of the Czech Republic
NS9678, research and development projectName: Biofilm - významný faktor virulence u infekcí močových katétrů; detekce a možnosti ovlivnění
Investor: Ministry of Health of the CR
VG20112015021, research and development projectName: Vývoj instrumentálních metodických postupů rychlé detekce a identifikace biologických agens v reálných vzorcích (Acronym: Detekce a identifikace biol. agens)
Investor: Ministry of the Interior of the CR
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