Identifying inhibitors/enhancers of quantitative real-time PCR in food samples using a newly developed synthetic plasmid
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027006%3A_____%2F16%3A00003433" target="_blank" >RIV/00027006:_____/16:00003433 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1002/jsfa.7178" target="_blank" >http://dx.doi.org/10.1002/jsfa.7178</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1002/jsfa.7178" target="_blank" >10.1002/jsfa.7178</a>
Alternative languages
Result language
angličtina
Original language name
Identifying inhibitors/enhancers of quantitative real-time PCR in food samples using a newly developed synthetic plasmid
Original language description
Polymerase chain reaction (PCR) has become a common technique offering fast and sensitive analysis of DNA in food/feed samples. However, many substances, either already present in the sample or introduced during sample processing, inhibit PCR and thus underestimate the DNA content. It is therefore necessary to identify PCR inhibition in order to correctly evaluate the sample. We designed and validated a synthetic plasmid DNA that can be used to detect and quantify PCR inhibition. The DNA sequence, appropriate primers and probe, were designed in silico, synthesized and the sequence was inserted into a plasmid vector. The performance of the plasmid was verified via calibration curves and by performing the assay in the presence of various DNAs (crops, fungus, bacterium). The detection of PCR inhibition was assessed using six inhibiting substances with different modes of action, substances used in sample processing (EDTA, ethanol, NaCl, SDS) and food additives (sodium glutamate, tartrazine). The plasmid performance proved to be reproducible and there were no interactions with other DNAs. The plasmid was able to identify the presence of the inhibitors in a wide range of concentrations. The presented plasmid DNA is a suitable and inexpensive possibility for evaluating PCR inhibition.
Czech name
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Czech description
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Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
GM - Food industry
OECD FORD branch
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Result continuities
Project
<a href="/en/project/QI101B267" target="_blank" >QI101B267: Development and application of effective procedures for control food production quality and their safety for consumer</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2016
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of the Science of Food and Agriculture
ISSN
0022-5142
e-ISSN
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Volume of the periodical
96
Issue of the periodical within the volume
3
Country of publishing house
GB - UNITED KINGDOM
Number of pages
5
Pages from-to
997-1001
UT code for WoS article
000368850500036
EID of the result in the Scopus database
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