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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

  • Czech description

Classification

  • Type

    J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

  • CEP classification

    GM - Food industry

  • OECD FORD branch

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

  • 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