TITLE

A preamplification approach to GMO detection in processed foods

AUTHOR(S)
Del Gaudio, S.; Cirillo, A.; Di Bernardo, G.; Galderisi, U.; Cipollaro, M.
PUB. DATE
March 2010
SOURCE
Analytical & Bioanalytical Chemistry;Mar2010, Vol. 396 Issue 6, p2135
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
DNA is widely used as a target for GMO analysis because of its stability and high detectability. Real-time PCR is the method routinely used in most analytical laboratories due to its quantitative performance and great sensitivity. Accurate DNA detection and quantification is dependent on the specificity and sensitivity of the amplification protocol as well as on the quality and quantity of the DNA used in the PCR reaction. In order to enhance the sensitivity of real-time PCR and consequently expand the number of analyzable target genes, we applied a preamplification technique to processed foods where DNA can be present in low amounts and/or in degraded forms thereby affecting the reliability of qualitative and quantitative results. The preamplification procedure utilizes a pool of primers targeting genes of interest and is followed by real-time PCR reactions specific for each gene. An improvement of Ct values was found comparing preamplified vs. non-preamplified DNA. The strategy reported in the present study will be also applicable to other fields requiring quantitative DNA testing by real-time PCR.
ACCESSION #
48536686

 

Related Articles

  • Quantification of relative flying fish paste content in the processed seafood ago- noyaki using real-time PCR. Nagase, Mitsutoshi; Yi, Ruirong; Hidaka, Fuminori; Maeta, Kazuhiko; Aimi, Tadanori; Yamaguchi, Takeshi; Suginaka, Katsuaki; Morinaga, Tsutomu // Fisheries Science;Sep2010, Vol. 76 Issue 5, p885 

    Real-time polymerase chain reaction (PCR) analysis of the 3′-portion of the mitochondrial 16S RNA gene (rDNA) coding sequence was used to determine flying fish paste in ago- noyaki. We quantified the amount of flying fish paste in ago- noyaki samples using flying fish-specific primers...

  • A statistical approach to quantification of genetically modified organisms (GMO) using frequency distributions. Gerdes, Lars; Busch, Ulrich; Pecoraro, Sven // BMC Bioinformatics;2014, Vol. 15 Issue 1, p593 

    Background According to Regulation (EU) No 619/2011, trace amounts of non-authorised genetically modified organisms (GMO) in feed are tolerated within the EU if certain prerequisites are met. Tolerable traces must not exceed the so-called 'minimum required performance limit' (MRPL), which was...

  • Assessment of the genetic stability of GMOs with a detailed examination of MON810 using Scorpion probes. Neumann, Georg; Brandes, Christian; Joachimsthaler, Alexandra; Hochegger, Rupert // European Food Research & Technology;Jul2011, Vol. 233 Issue 1, p19 

    For the cultivation and maintenance of seeds, genetic mutations have to be avoided as much as possible. According to the Directive 2001/18/EC, genetic stability has to be proven for the approval of GMOs []. The investigation of genetic stability can be performed with various methods, but...

  • Detection of dog and cat traces in food, pet food and farm animal feed by real-time PCR. Espiñeira, Montserrat; Vieites, Juan // European Food Research & Technology;Aug2015, Vol. 241 Issue 2, p233 

    The identification of species presents in food is important due to many reasons. Some of them are economic fraud and/or sanitary problems, which could arise from inaccurate identifications of the composition of food products. Adulteration of processed meat generally occurs when there is a...

  • ASPECTS REGARDING THE PROFILE OF INTESTINAL MICROBIOTA ON WILD POPULATIONS OF STERLET (ACIPENSER RUTHENUS, LINNAEUS, 1758). BĂCANU, MANUELA GIANINA; OPREA, LUCIAN; SANDU, GEORGIANA CĂLIN; DINICĂ, RODICA; MĂEREANU, MARILENA; MĂEREANU, DUMITRU; ŞERBAN, CONSTANŢA // Annals of the University Dunarea de Jos of Galati Fascicle VI --;2012, Vol. 36 Issue 2, p58 

    The aim of the present research was to examinate the profile of intestinal microbiota of sterlet from the Danube River. Genomic DNA was extracted from each gut fish and polymerase chain reaction (PCR) was used to amplify the conserved 16S ribosomal RNA gene. Using Denaturing Gradient Gel...

  • Optimization of PCR in application of hot start Taq DNA polymerase for detection of Erwinia amylovora with primers FER1-F and FER1-R1. Obradovic, D.; Kevresan, S. // Microbiology (00262617);Dec2010, Vol. 79 Issue 6, p816 

    There are two approaches in detection of bacterium Erwinia amylovora by PCR. One is based on detection of plasmid pEA29 and the other is based on detection of a chromosomal DNA sequence, specific for E. amylovora, in a sample. Since pathogenic strains without pEA29 have been isolated from the...

  • Quantitative polymerase chain reaction analysis by deconvolution of internal standard. Hirakawa, Yasuko; Medh, Rheem D.; Metzenberg, Stan // BMC Molecular Biology;2010, Vol. 11, p30 

    Background: Quantitative Polymerase Chain Reaction (qPCR) is a collection of methods for estimating the number of copies of a specific DNA template in a sample, but one that is not universally accepted because it can lead to highly inaccurate (albeit precise) results. The fundamental problem is...

  • Characterization of a Methanogenic Community within an Algal Fed Anaerobic Digester. Ellis, Joshua T.; Tramp, Cody; Sims, Ronald C.; Miller, Charles D. // ISRN Microbiology;2012, p1 

    The article presents information on a study conducted to characterize the methanogenic community within an algal fed anaerobic digester. It informs that the during the study the DNA was extracted from anaerobic sludge material and used in metagenomic analysis through polymerase chain reaction...

  • Implementation of Novel Design Features for qPCR-Based eDNA Assessment. Veldhoen, Nik; Hobbs, Jared; Ikonomou, Georgios; Hii, Michael; Lesperance, Mary; Helbing, Caren C. // PLoS ONE;11/1/2016, Vol. 11 Issue 10, p1 

    Environmental stewardship requires timely, accurate information related to the status of a given ecosystem and the species that occupy it. Recent advances in the application of the highly sensitive real-time quantitative polymerase chain reaction (qPCR) towards identification of constituents...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics