Electrochemical assay for sensitive detection of nucleic acid of african swine fever virus

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43510%2F19%3A43920908" target="_blank" >RIV/62156489:43510/19:43920908 - isvavai.cz</a>

Výsledek na webu

<a href="http://www.biocap.ma/wp-content/uploads/2019/10/Book-of-abstract.pdf" target="_blank" >http://www.biocap.ma/wp-content/uploads/2019/10/Book-of-abstract.pdf</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Electrochemical assay for sensitive detection of nucleic acid of african swine fever virus

Popis výsledku v původním jazyce

Introduction. African Swine Fever Virus (ASFV) is a DNA virus of the Asfivirus genus of the Asfarviridae family that is found in blood, body fluids, and internal organs1,2. ASFV was described more than 40 years ago3,4. This virus spreads pandemically and the mortality rate of the virus-related disease ranges from 90 to 100 %5,6. The aim of this study was to propose the detection of specific nucleic acid (NA) of ASFV using electrochemical hybridization biosensor. Experimental part. The absorbance spectra of nanoparticles were measured in a UV-VIS spectrophotometer. Determination of DNA and/or RNA by DPV and/or SWV was performed using Autolab. The parameters of the measurement were as follows: initial potential 0 V, end potential -1.8 V, accumulation time 120 s, step potential 5 mV, modulation amplitude 25 mV, frequency 200-500 Hz, the volume of injected sample 10 μL. Results and discussion. Electrochemical methods are suitable techniques for the study of ODNs, PCR products and their components. The AdTS detection limit of NA samples (signals of adenine and cytosine around -1.3 V) was about hundreds of attomoles per 5 μL drop. To increase the selectivity and sensitivity of NA determination, we have prepared electrochemical labels (CdTe) by green synthesis7. Using these labels, specific probes for the target DNA were labelled. A meat product (ham) was used as a sample. After DNA extraction, PCR reaction to the target sequence was performed. The PCR product was subsequently recognized by the probe labelled with CdTe. The proposed procedure has increased the sensitivity of PCR product detection by 85 %. Conclusion. We have designed a sensitive electrochemical method for detecting viral nucleotide sequence. The detection hybridization probe used was labelled with green synthesis-prepared CdTe.

Název v anglickém jazyce

Electrochemical assay for sensitive detection of nucleic acid of african swine fever virus

Popis výsledku anglicky

Introduction. African Swine Fever Virus (ASFV) is a DNA virus of the Asfivirus genus of the Asfarviridae family that is found in blood, body fluids, and internal organs1,2. ASFV was described more than 40 years ago3,4. This virus spreads pandemically and the mortality rate of the virus-related disease ranges from 90 to 100 %5,6. The aim of this study was to propose the detection of specific nucleic acid (NA) of ASFV using electrochemical hybridization biosensor. Experimental part. The absorbance spectra of nanoparticles were measured in a UV-VIS spectrophotometer. Determination of DNA and/or RNA by DPV and/or SWV was performed using Autolab. The parameters of the measurement were as follows: initial potential 0 V, end potential -1.8 V, accumulation time 120 s, step potential 5 mV, modulation amplitude 25 mV, frequency 200-500 Hz, the volume of injected sample 10 μL. Results and discussion. Electrochemical methods are suitable techniques for the study of ODNs, PCR products and their components. The AdTS detection limit of NA samples (signals of adenine and cytosine around -1.3 V) was about hundreds of attomoles per 5 μL drop. To increase the selectivity and sensitivity of NA determination, we have prepared electrochemical labels (CdTe) by green synthesis7. Using these labels, specific probes for the target DNA were labelled. A meat product (ham) was used as a sample. After DNA extraction, PCR reaction to the target sequence was performed. The PCR product was subsequently recognized by the probe labelled with CdTe. The proposed procedure has increased the sensitivity of PCR product detection by 85 %. Conclusion. We have designed a sensitive electrochemical method for detecting viral nucleotide sequence. The detection hybridization probe used was labelled with green synthesis-prepared CdTe.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10405 - Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis)

Projekt

<a href="/cs/project/LTC18002" target="_blank" >LTC18002: Vývoj nových materiálů vhodných pro 3D tisk s antimikrobiálními vlastnostmi (3D ANTIMICROB)</a><br>

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