Portable immunosensor systems for detection of Francisella
tularensis

D 2006

Portable immunosensor systems for detection of Francisella tularensis

SKLÁDAL, Petr; Miroslav POHANKA; Jan PŘIBYL; Milan JÍLEK; Bohuslav ŠAFÁŘ et al.

Základní údaje

Originální název

Portable immunosensor systems for detection of Francisella tularensis

Název česky

Přenosné imunosensorové systémy pro detekci Francisella tularensis

Autoři

SKLÁDAL, Petr; Miroslav POHANKA; Jan PŘIBYL ; Milan JÍLEK a Bohuslav ŠAFÁŘ

Vydání

Brno, Commercial and Pre-commercial Cell Detection Technologies for Defence against Bioterror – Technology, Market and Society. NATO Advanced Research Workshop, s. 17-17, 2006

Nakladatel

Masarykova universita

Další údaje

Jazyk

angličtina

Typ výsledku

Stať ve sborníku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

Amperometric immunosensor; Screen printed electrode; Portable detector

Štítky

Amperometric immunosensor, Portable detector, Screen printed electrode

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 13. 1. 2007 11:10, prof. RNDr. Petr Skládal, CSc.

Anotace

ORIG CZ

V originále

The electrochemical sensors and piezoelectric resonators were adopted as two alternatives of robust, sensitive and affordable transducers suitable for construction of portable detection systems. The former approach represents a highly-sensitive immunospecific detection of bioagents using tracers based on the peroxidase enzyme label (both sandwich and competitive assay formats) providing electrochemically active products measured using the screen-printed electrochemical systems. In this way, the biosensing part of the detector is easily exchanged after positive detection and disposable use is preferred. However, the heterogeneous sensing format requires washing steps and several working solutions, which are delivered by the embedded miniperistaltic pumps. The piezoelectric biosensor provides direct measurement of the target bioagents, the sensing part is used repeatedly after regeneration of the immobilized antibody. From the construction point of view, both lines of detectors share several common elements (case, rechargeable battery, microcontroller, A/D converters, pumps), though the sensing parts are different: multipotentiostat / oscillator and counter, custom flow-through cells and biosensors. Detectors are linked to the external computer with software LabTools (common data-management and user interface, individual detector-related plugin modules). For testing purposes, the model bioagent was the bacterium Francisella tularensis (Ft) included on the lists of biowarfare agents (the LVS variant was employed), several other safe bioagent simulants were recently included, too. For immobilization of biorecognition elements, the gold-coated sensing surface of either working electrode or quartz crystal resonator was modified with a self-assembled monolayer of thiocompound which was covalently linked to the specific proteins - antibodies and antigens as lipopolysaccharide fraction (LPS). The electrochemical detector was able to detect F. tularensis starting at 100 CFU/ml, one analysis (incubations with sample and tracer, washing, addition of sample, amperometric measurement) was completed within 15 min. The piezoelectric detector with immobilized specific antibodies was suitable to indicate the presence of 105 CFU/ml F. tularensis directly within 10 min, the response of 4.8 Hz was significantly higher compared to non-specific species (108 CFU/ml E. coli and B. subtilis provided signals below 2 Hz). In the case of the piezoelectric immunosensor modified with LPS, rapid detection of anti-Ft antibodies in mice inoculated with the bioagent (0.1 LD50) was possible, specific response was obtained shortly (1-3 days) after infection. In this way, the piezoelectric immunosensor serves as suitable tool for sensitive detection of tularemic infection before appearance of any clinical disease symptoms. The developed biosensor-based devices are promising tools suitable for rapid detection of bioagents as well as other biomolecules outside of laboratory. The piezoelectric detector is already commercialized, the electrochemical detector is at the phase of realization of field tests using model agents as B. subtilis, thuringiensis and Erwinia herbicola.

Česky

Byl vyvinut amperometrický multikanálový imunosensor na bázi sítotiskového elektrochemického systému. Tento sensor je začleněn do přenosného biosensorového detektoru MultiLab. Testování probíhá na různých typech mikroorganismů, včetně bakterie Francisella tularensis, která je potenciálně zneužitelná bioteroristy.

Návaznosti

LC06030, projekt VaV

Název: Biomolekulární centrum

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Biomolekulární centrum

Citovat

SKLÁDAL, Petr; Miroslav POHANKA; Jan PŘIBYL; Milan JÍLEK a Bohuslav ŠAFÁŘ. Portable immunosensor systems for detection of Francisella tularensis. In Commercial and Pre-commercial Cell Detection Technologies for Defence against Bioterror – Technology, Market and Society. NATO Advanced Research Workshop. Brno: Masarykova universita, 2006, s. 17.

@inproceedings{708778,
   author = {Skládal, Petr and Pohanka, Miroslav and Přibyl, Jan and Jílek, Milan and Šafář, Bohuslav},
   address = {Brno},
   booktitle = {Commercial and Pre-commercial Cell Detection Technologies for Defence against Bioterror – Technology, Market and Society. NATO Advanced Research Workshop},
   keywords = {Amperometric immunosensor; Screen printed electrode; Portable detector},
   language = {eng},
   location = {Brno},
   pages = {17-17},
   publisher = {Masarykova universita},
   title = {Portable immunosensor systems for detection of Francisella tularensis},
   year = {2006}
}

TY  - CONF
ID  - 708778
AU  - Skládal, Petr - Pohanka, Miroslav - Přibyl, Jan - Jílek, Milan - Šafář, Bohuslav
PY  - 2006
TI  - Portable immunosensor systems for detection of Francisella tularensis
PB  - Masarykova universita
CY  - Brno
KW  - Amperometric immunosensor
KW  - Screen printed electrode
KW  - Portable detector
N2  - The electrochemical sensors and piezoelectric resonators were adopted as two alternatives of robust, sensitive and affordable transducers suitable for construction of portable detection systems. The former approach represents a highly-sensitive immunospecific detection of bioagents using tracers based on the peroxidase enzyme label (both sandwich and competitive assay formats) providing electrochemically active products measured using the screen-printed electrochemical systems. In this way, the biosensing part of the detector is easily exchanged after positive detection and disposable use is preferred. However, the heterogeneous sensing format requires washing steps and several working solutions, which are delivered by the embedded miniperistaltic pumps. The piezoelectric biosensor provides direct measurement of the target bioagents, the sensing part is used repeatedly after regeneration of the immobilized antibody. From the construction point of view, both lines of detectors share several common elements (case, rechargeable battery, microcontroller, A/D converters, pumps), though the sensing parts are different: multipotentiostat / oscillator and counter, custom flow-through cells and biosensors. Detectors are linked to the external computer with software LabTools (common data-management and user interface, individual detector-related plugin modules). For testing purposes, the model bioagent was the bacterium Francisella tularensis (Ft) included on the lists of biowarfare agents (the LVS variant was employed), several other safe bioagent simulants were recently included, too. For immobilization of biorecognition elements, the gold-coated sensing surface of either working electrode or quartz crystal resonator was modified with a self-assembled monolayer of thiocompound which was covalently linked to the specific proteins - antibodies and antigens as lipopolysaccharide fraction (LPS). The electrochemical detector was able to detect F. tularensis starting at 100 CFU/ml, one analysis (incubations with sample and tracer, washing, addition of sample, amperometric measurement) was completed within 15 min. The piezoelectric detector with immobilized specific antibodies was suitable to indicate the presence of 105 CFU/ml F. tularensis directly within 10 min, the response of 4.8 Hz was significantly higher compared to non-specific species (108 CFU/ml E. coli and B. subtilis provided signals below 2 Hz). In the case of the piezoelectric immunosensor modified with LPS, rapid detection of anti-Ft antibodies in mice inoculated with the bioagent (0.1 LD50) was possible, specific response was obtained shortly (1-3 days) after infection. In this way, the piezoelectric immunosensor serves as suitable tool for sensitive detection of tularemic infection before appearance of any clinical disease symptoms. The developed biosensor-based devices are promising tools suitable for rapid detection of bioagents as well as other biomolecules outside of laboratory. The piezoelectric detector is already commercialized, the electrochemical detector is at the phase of realization of field tests using model agents as B. subtilis, thuringiensis and Erwinia herbicola.
ER  -

SKLÁDAL, Petr; Miroslav POHANKA; Jan PŘIBYL; Milan JÍLEK a Bohuslav ŠAFÁŘ. Portable immunosensor systems for detection of Francisella tularensis. In \textit{Commercial and Pre-commercial Cell Detection Technologies for Defence against Bioterror – Technology, Market and Society. NATO Advanced Research Workshop}. Brno: Masarykova universita, 2006, s.~17.

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