Detection of mandarin in orange juice by single-nucleotide polymorphism qPCR assay
Identifieur interne : 000C60 ( Main/Exploration ); précédent : 000C59; suivant : 000C61Detection of mandarin in orange juice by single-nucleotide polymorphism qPCR assay
Auteurs : Miriam Aldeguer [Espagne] ; Maria Lopez-Andreo [Espagne] ; José A. Gabaldon [Espagne] ; Antonio Puyet [Espagne]Source :
- Food chemistry [ 0308-8146 ] ; 2014.
Descripteurs français
- Pascal (Inist)
- Wicri :
- geographic : Espagne.
English descriptors
- KwdEn :
- Adulteration, Assay, Beverages (analysis), Citrus, Citrus (chemistry), Citrus (genetics), Citrus (metabolism), Citrus sinensis (chemistry), Crosses, Genetic, DNA Copy Number Variations, DNA Probes (analysis), DNA Probes (chemistry), DNA Probes (metabolism), DNA, Intergenic (analysis), DNA, Intergenic (chemistry), DNA, Intergenic (metabolism), DNA, Plant (analysis), DNA, Plant (chemistry), DNA, Plant (metabolism), Detection, Fluorescent Dyes (chemistry), Food, Food Contamination, Food Handling, Food Inspection (methods), Food Labeling, Food Quality, Fruit (chemistry), Fruit (genetics), Fruit (metabolism), Identification, Limit of Detection, Nucleotide, Orange juice, Polymerase chain reaction, Polymorphism, Polymorphism, Single Nucleotide, Probe, Quality, Real time, Real-Time Polymerase Chain Reaction, Rhodamines (chemistry), Spain, Species Specificity.
- MESH :
- chemical , analysis : DNA Probes, DNA, Intergenic, DNA, Plant.
- geographic : Spain.
- analysis : Beverages.
- chemistry : Citrus, Citrus sinensis, DNA Probes, DNA, Intergenic, DNA, Plant, Fluorescent Dyes, Fruit, Rhodamines.
- genetics : Citrus, Fruit.
- metabolism : Citrus, DNA Probes, DNA, Intergenic, DNA, Plant, Fruit.
- methods : Food Inspection.
- Crosses, Genetic, DNA Copy Number Variations, Food Contamination, Food Handling, Food Labeling, Food Quality, Limit of Detection, Polymorphism, Single Nucleotide, Real-Time Polymerase Chain Reaction, Species Specificity.
Abstract
A dual-probe real time PCR (qPCR) DNA-based analysis was devised for the identification of mandarin in orange juice. A single nucleotide polymorphism at the trnL-trnF intergenic region of the chloroplast chromosome was confirmed in nine orange (Citrus sinensis) and thirteen commercial varieties of mandarin, including Citrus reticulata and Citrus unshiu species and a mandarin x tangelo hybrid. Two short minor-groove binding fluorescent probes targeting the polymorphic sequence were used in the dual-probe qPCR, which allowed the detection of both species in single-tube reactions. The similarity of PCR efficiencies allowed a simple estimation of the ratio mandarin/orange in the juice samples, which correlated to the measured difference of threshold cycle values for both probes. The limit of detection of the assay was 5% of mandarin in orange juice, both when the juice was freshly prepared (not from concentrate) or reconstituted from concentrate, which would allow the detection of fraudulently added mandarin juice. The possible use of the dual-probe system for quantitative measurements was also tested on fruit juice mixtures. qPCR data obtained from samples containing equal amounts of mandarin and orange juice revealed that the mandarin target copy number was approximately 2.6-fold higher than in orange juice. The use of a matrix-adapted control as calibrator to compensate the resulting CT bias allowed accurate quantitative measurements to be obtained.
Affiliations:
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Le document en format XML
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Gabaldon</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Centro Tecnológico Nacional de la Conserva</s1><s2>Molina de Segura, Murcia</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Espagne</country><wicri:noRegion>Centro Tecnológico Nacional de la Conserva</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Departamento de Ciencia y Tecnología de Alimentos, Universidad Católica San Antonio de Murcia (UCAM)</s1><s2>Murcia</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Espagne</country><wicri:noRegion>Murcia</wicri:noRegion></affiliation></author><author><name sortKey="Puyet, Antonio" sort="Puyet, Antonio" uniqKey="Puyet A" first="Antonio" last="Puyet">Antonio Puyet</name><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Departamento de Bioquímica y Biologia Molecular IV, Facultad de Veterinaria, Universidad Complutense de Madrid</s1><s2>28040 Madrid</s2><s3>ESP</s3><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>Espagne</country><placeName><region nuts="2" type="communauté">Communauté de Madrid</region><settlement type="city">Madrid</settlement></placeName><orgName type="university">Université complutense de Madrid</orgName></affiliation></author></analytic><series><title level="j" type="main">Food chemistry</title><title level="j" type="abbreviated">Food chem.</title><idno type="ISSN">0308-8146</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Food chemistry</title><title level="j" type="abbreviated">Food chem.</title><idno type="ISSN">0308-8146</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adulteration</term><term>Assay</term><term>Beverages (analysis)</term><term>Citrus</term><term>Citrus (chemistry)</term><term>Citrus (genetics)</term><term>Citrus (metabolism)</term><term>Citrus sinensis (chemistry)</term><term>Crosses, Genetic</term><term>DNA Copy Number Variations</term><term>DNA Probes (analysis)</term><term>DNA Probes (chemistry)</term><term>DNA Probes (metabolism)</term><term>DNA, Intergenic (analysis)</term><term>DNA, Intergenic (chemistry)</term><term>DNA, Intergenic (metabolism)</term><term>DNA, Plant (analysis)</term><term>DNA, Plant (chemistry)</term><term>DNA, Plant (metabolism)</term><term>Detection</term><term>Fluorescent Dyes (chemistry)</term><term>Food</term><term>Food Contamination</term><term>Food Handling</term><term>Food Inspection (methods)</term><term>Food Labeling</term><term>Food Quality</term><term>Fruit (chemistry)</term><term>Fruit (genetics)</term><term>Fruit (metabolism)</term><term>Identification</term><term>Limit of Detection</term><term>Nucleotide</term><term>Orange juice</term><term>Polymerase chain reaction</term><term>Polymorphism</term><term>Polymorphism, Single Nucleotide</term><term>Probe</term><term>Quality</term><term>Real time</term><term>Real-Time Polymerase Chain Reaction</term><term>Rhodamines (chemistry)</term><term>Spain</term><term>Species Specificity</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>DNA Probes</term><term>DNA, Intergenic</term><term>DNA, Plant</term></keywords><keywords scheme="MESH" type="geographic" xml:lang="en"><term>Spain</term></keywords><keywords scheme="MESH" qualifier="analysis" xml:lang="en"><term>Beverages</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Citrus</term><term>Citrus sinensis</term><term>DNA Probes</term><term>DNA, Intergenic</term><term>DNA, Plant</term><term>Fluorescent Dyes</term><term>Fruit</term><term>Rhodamines</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Citrus</term><term>Fruit</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Citrus</term><term>DNA Probes</term><term>DNA, Intergenic</term><term>DNA, Plant</term><term>Fruit</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Food Inspection</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Crosses, Genetic</term><term>DNA Copy Number Variations</term><term>Food Contamination</term><term>Food Handling</term><term>Food Labeling</term><term>Food Quality</term><term>Limit of Detection</term><term>Polymorphism, Single Nucleotide</term><term>Real-Time Polymerase Chain Reaction</term><term>Species Specificity</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Détection</term><term>Jus d'orange</term><term>Nucléotide</term><term>Polymorphisme</term><term>Dosage</term><term>Identification</term><term>Aliment</term><term>Qualité</term><term>Temps réel</term><term>Réaction chaîne polymérase</term><term>Sonde</term><term>Adultération</term><term>Citrus</term></keywords><keywords scheme="Wicri" type="geographic" xml:lang="fr"><term>Espagne</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A dual-probe real time PCR (qPCR) DNA-based analysis was devised for the identification of mandarin in orange juice. A single nucleotide polymorphism at the trnL-trnF intergenic region of the chloroplast chromosome was confirmed in nine orange (Citrus sinensis) and thirteen commercial varieties of mandarin, including Citrus reticulata and Citrus unshiu species and a mandarin x tangelo hybrid. Two short minor-groove binding fluorescent probes targeting the polymorphic sequence were used in the dual-probe qPCR, which allowed the detection of both species in single-tube reactions. The similarity of PCR efficiencies allowed a simple estimation of the ratio mandarin/orange in the juice samples, which correlated to the measured difference of threshold cycle values for both probes. The limit of detection of the assay was 5% of mandarin in orange juice, both when the juice was freshly prepared (not from concentrate) or reconstituted from concentrate, which would allow the detection of fraudulently added mandarin juice. The possible use of the dual-probe system for quantitative measurements was also tested on fruit juice mixtures. qPCR data obtained from samples containing equal amounts of mandarin and orange juice revealed that the mandarin target copy number was approximately 2.6-fold higher than in orange juice. The use of a matrix-adapted control as calibrator to compensate the resulting C<sub>T</sub> bias allowed accurate quantitative measurements to be obtained.</div></front></TEI><affiliations><list><country><li>Espagne</li></country><region><li>Communauté de Madrid</li></region><settlement><li>Madrid</li></settlement><orgName><li>Université complutense de Madrid</li></orgName></list><tree><country name="Espagne"><noRegion><name sortKey="Aldeguer, Miriam" sort="Aldeguer, Miriam" uniqKey="Aldeguer M" first="Miriam" last="Aldeguer">Miriam Aldeguer</name></noRegion><name sortKey="Aldeguer, Miriam" sort="Aldeguer, Miriam" uniqKey="Aldeguer M" first="Miriam" last="Aldeguer">Miriam Aldeguer</name><name sortKey="Gabaldon, Jose A" sort="Gabaldon, Jose A" uniqKey="Gabaldon J" first="José A." last="Gabaldon">José A. Gabaldon</name><name sortKey="Gabaldon, Jose A" sort="Gabaldon, Jose A" uniqKey="Gabaldon J" first="José A." last="Gabaldon">José A. Gabaldon</name><name sortKey="Lopez Andreo, Maria" sort="Lopez Andreo, Maria" uniqKey="Lopez Andreo M" first="Maria" last="Lopez-Andreo">Maria Lopez-Andreo</name><name sortKey="Puyet, Antonio" sort="Puyet, Antonio" uniqKey="Puyet A" first="Antonio" last="Puyet">Antonio Puyet</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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