Quantitative competitive PCR for the detection of genetically modified soybean and maize

Studer, Edgar; Rhyner, Claudio; Lüthy, Jürg; Hübner, Philipp

doi:10.1007/s002170050320

Cited by 115 publications

(62 citation statements)

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“…Competitor was constructed similar to strategies described earlier (Grassi et al, 1994;Schanke et al, 1994;Studer et al, 1998). The competitor (external control) was designed to contain Pou-universal primer binding sequence of ~20 bp at ends encompassing a phage sequence.…”

Section: Construction Of the Poultry Dna Competitormentioning

confidence: 99%

Development and use of quantitative competitive PCR assay for detection of poultry DNA in fish meal

Farajollahi¹,

Aslaminejad²,

Nassiry³

et al. 2009

J. Anim. Feed Sci.

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In this study we reported the development and evaluation of a quantitative competitive polymerase chain reaction (QC-PCR) for detection and quantification of poultry DNA in fish meal. A universal PCR primer pair (Pou) was used to amplify a specific region 12s rRNA from poultry DNA. Specificity of the primers was evaluated with DNA from cattle and sheep. An external control DNA was constructed by 83 bp differing in length from the poultry target sequence and used for PCR reaction together with the target DNA. DNA was extracted from thirty commercial samples of fish meal and spiked with external control DNA and was co-amplified with Pou primer pair. The signal intensity was quantified by ImageJ 1.29x and were used to compare variety of poultry pollution percentage in fish meal samples. The results of QC_PCR showed that pollution percentage was in the range of 0.1 to 3% in collected samples. This study was the first report of QC_PCR application for quantifying of poultry pollution in fish meal in Iran.

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Section: Construction Of the Poultry Dna Competitormentioning

confidence: 99%

Development and use of quantitative competitive PCR assay for detection of poultry DNA in fish meal

Farajollahi¹,

Aslaminejad²,

Nassiry³

et al. 2009

J. Anim. Feed Sci.

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“…Nevertheless, the limited sensitivity and resolution, and the difficulties to get quantitative results and for automation, have forced the development of alternatives, either for the detection itself of both for separation and detection. Examples of alternative detection methods are the use of CCD cameras instead of photography, 32,45 or the use of isotopic labelling of the PCR products, followed by electrophoresis and detection by autoradiography or scintillation counting of gel slices. 46,47 Independently of the detection method employed, one major drawback of methods based in agarose or acrylamide gel electrophoresis is that two steps are necessary in order to perform the analysis, which extends the analysis time and hinders the accuracy and precision of the results.…”

Section: End-point Detection Of Pcr Productsmentioning

confidence: 99%

“…In these procedures, PCR products are analysed by conventional agarose gel electrophoresis, ethidium bromide staining and visual estimation of the equivalence point 65 or computer analysis of a digital image obtained with a CCD camera. 32,45 Certified reference materials are commonly used for calibration of these systems, but, as mentioned above, this could lead to underestimation of specific transgenic varieties in processed food. 66 In order to overcome this problem, quantification results for a transgenic sequence are referred to the quantity of a control target sequence, preferentially of a similar size, that should be present in equal amounts in the transgenic and the conventional varieties.…”

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confidence: 99%

Detection of Genetically Modified Organisms in Foods by DNA Amplification Techniques

García‐Cañas

Cifuentes

González

2004

Critical Reviews in Food Science and Nutrition

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“…Certaines sont basées sur l'analyse immunochimique (ELISA) et nécessitent l'utilisation d'anticorps monoclonaux spécifiques de la protéine codée par le transgène [8,9]. D'autres reposent sur la PCR quantitative compétitive [10] ou la PCR quantitative en temps réel [11][12][13].…”

Section: Introductionunclassified

La PCR quantitative en temps réel : application à la quantification des OGM

Alary

Gautier

Joudrier

2002

OCL

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Résumé : Suite à l'obligation d'étiquetage, au seuil de 1 %, des aliments contenant des OGM autorisés, il est nécessaire de disposer de méthodes fiables de quantification. Pour répondre à cette obligation, la technique de PCR quantitative en temps réel semble actuellement la mieux adaptée. Son principe, ses avantages et sa mise en oeuvre pour la détermination de la teneur en OGM de farines de soja sont présentés. Les PCR simplex et duplex sont comparées. Summary :Because of the GMO food labelling requirement at the threshold of 1%, the availability of reliable quantitative analytical methods is necessary. The real-time quantitative PCR seems to be the most accurate method for GMO quantification. Its principle, advantages and application to the determination of GMO content of soybean flour samples are presented. Simplex and duplex PCRs are compared. Mots-clés :OGM, soja, PCR quantitative en temps réel, ADN, étiquetage.

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Quantitative competitive PCR for the detection of genetically modified soybean and maize

Cited by 115 publications

References 0 publications

Development and use of quantitative competitive PCR assay for detection of poultry DNA in fish meal

Development and use of quantitative competitive PCR assay for detection of poultry DNA in fish meal

Detection of Genetically Modified Organisms in Foods by DNA Amplification Techniques

La PCR quantitative en temps réel : application à la quantification des OGM

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