Development of detection panel for the simultaneous screening of biothreat agents - B. anthracis, Y. pestis, F. tularensis and Brucella spp. by MOL-PCR method

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F19%3AN0000280" target="_blank" >RIV/00027162:_____/19:N0000280 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00027162:_____/19:N0000325

Výsledek na webu

—

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

Development of detection panel for the simultaneous screening of biothreat agents - B. anthracis, Y. pestis, F. tularensis and Brucella spp. by MOL-PCR method

Popis výsledku v původním jazyce

xMAP CONNECT, 2019 5th ANNIVERSARY, AMSTERDAM, 5. – 6.11. 2019 - poster. Molecular biological methods, such as polymerase chain reaction (PCR), are very important for the rapid detection of pathogenic agents in the human and veterinary field. Early detection is important for the treatment of infectious diseases. Currently, a large number of pathogens need to be identified in a short time in a single reaction. We have developed and validated detection panel based on multiplex method based on oligonucleotide ligation followed by PCR for bacteria that can be used as a biological weapon. The panel detects bacteria, which are classified in category A - most dangerous (B. anthracis, Y. pestis, F. tularensis) and B - less dangerous pathogens (Brucella spp.). For this reason, we used xMAP technology (x = analyte, MAP = Multi Analyte Profiling) using magnetic microspheres to detect nucleic acids (specific genes). Multiplex oligonucleotide ligation (MOL) followed by PCR with fluorescently labelled primer was optimized in this protocol. The final step was the hybridization of PCR products to magnetic microspheres and MagPix analysis. Finally, the designed oligonucleotide probes for detection of selected bacteria were tested and the detection panel was validated to fulfil the requirements for the routine diagnostic method. This includes determination of specificity and sensitivity.

Název v anglickém jazyce

Development of detection panel for the simultaneous screening of biothreat agents - B. anthracis, Y. pestis, F. tularensis and Brucella spp. by MOL-PCR method

Popis výsledku anglicky

xMAP CONNECT, 2019 5th ANNIVERSARY, AMSTERDAM, 5. – 6.11. 2019 - poster. Molecular biological methods, such as polymerase chain reaction (PCR), are very important for the rapid detection of pathogenic agents in the human and veterinary field. Early detection is important for the treatment of infectious diseases. Currently, a large number of pathogens need to be identified in a short time in a single reaction. We have developed and validated detection panel based on multiplex method based on oligonucleotide ligation followed by PCR for bacteria that can be used as a biological weapon. The panel detects bacteria, which are classified in category A - most dangerous (B. anthracis, Y. pestis, F. tularensis) and B - less dangerous pathogens (Brucella spp.). For this reason, we used xMAP technology (x = analyte, MAP = Multi Analyte Profiling) using magnetic microspheres to detect nucleic acids (specific genes). Multiplex oligonucleotide ligation (MOL) followed by PCR with fluorescently labelled primer was optimized in this protocol. The final step was the hybridization of PCR products to magnetic microspheres and MagPix analysis. Finally, the designed oligonucleotide probes for detection of selected bacteria were tested and the detection panel was validated to fulfil the requirements for the routine diagnostic method. This includes determination of specificity and sensitivity.

Druh

O - Ostatní výsledky

CEP obor

—

OECD FORD obor

10607 - Virology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů