A multiplex PCR assay for fraud identification of deer products

Zha, Daiming; Xing, Xiumei; Fu-he, Yang

doi:10.1016/j.foodcont.2010.04.013

Cited by 33 publications

(16 citation statements)

References 15 publications

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“…The limit of detection for the different methods is comparable to that obtained in another study using the same target (Lopéz-Andreo et al, 2005), lower than those obtained by Zha, Xing, and Yang (2010) using mitochondrial genes in an "end point" PCR and lower than those showed by Laube et al (2007).…”

Section: Discussionsupporting

confidence: 81%

Development and validation of fast Real-Time PCR assays for species identification in raw and cooked meat mixtures

2012

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Section: Discussionsupporting

confidence: 81%

Development and validation of fast Real-Time PCR assays for species identification in raw and cooked meat mixtures

2012

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“…This technique has also been applied to authenticate several other species, including sea cucumbers (Zuo et al, 2012) and mackerel (Catanese et al, 2010), and to uncover meat scandals, such as venison (Zha et al, 2010) and abalone products (Chan et al, 2012). Therefore, multiplex PCR with a novel primer set (KUGEN-ILSspec) could be an effective tool for prevention Downloaded by [University of Windsor] at 18:07 19 November 2014…”

Section: Pcr Amplificationmentioning

confidence: 99%

“…Such a low concentration of DNA templates was also adequate for detecting mixed templates of L. lunaris, L. spadiceus and L. inermis. The detection limit of 1 ng/µL concentration of DNA templates is considered as highly sensitive when compared to bovine-specific primers, poultry-specific primers (Zha et al, 2010), and Scomber-specific primers (Catanese et al, 2010). Therefore, our puffer fish specific primers could be effective for managing food fraud and safety control.…”

Section: Sensitivity Test Of Multiplex Pcr Primermentioning

confidence: 99%

Identification of Puffer Fish of the GenusLagocephalus: L. lunaris, L. spadiceusandL. inermis, Using Multiplex PCR

2014

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A multiplex PCR assay was developed to identify widespread poison-containing puffer fishes of the genus Lagocephalus: L. lunaris, L. spadiceus and L. inermis, using a novel KUGEN_ILSspec primer set which is specific for 12S ribosomal RNA (12S rRNA) and Cytochrome b (Cyt b) genes. KUGEN_ILSspec set is composed of fish positive-amplified primers that produced a 289-bp fragment while L. lunaris-, L. spadiceus-and L. inermis-specific primers produced 123-, 196-and 493-bp fragments, respectively. The sensitivity of this primer set was found to be high as it was capable of amplifying targeted DNA at concentrations as low as 1 ng/µL. Moreover, the primer set was shown to amplify intact DNA and species-specific fragments from heat-processed food. Consequently, multiplex PCR technique in combination with KUGEN_ILSspec primer set could offer an effective measure for detecting poisonous puffer fish contamination in both fresh and processed fish products, which will greatly aid in public health control and law enforcing endeavors.

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“…Since mtDNA occurs in high copy numbers in each cell and is able to withstand degradation and environmental challenges, it is well suited for use in food authentication assays. MtDNA assays have already been developed and used for species testing in food safety management and fraudulence using both end-point PCR and qPCR techniques (Dooley et al 2004;Köppel et al 2009;Woolfe and Primrose 2004;Dalmasso et al 2004;Zha et al 2010;Rodriguez et al 2003;Fumiere et al 2006;Eugster et al 2009;Tanabe et al 2007). Despite mtDNA's durability (Hajibabaei et al 2007), nuclear loci are more advantageous for DNA quantification because the diploid (instead of multiple) copy number Walker et al 2004) makes it more predictive of nuclear DNA profiling success and more relevant to identity testing and sample traceability than multi-copy and/or non-nuclear markers (Alonso et al 2004;Timken et al 2005).…”

Section: Introductionmentioning

confidence: 99%

“…The integration of species determination and nuclear DNA quantification into a single assay will streamline downstream genotyping analysis, and improve the quality of the DNA profiles while conserving reagents (Evans et al 2007;Lindquist et al 2011;Kanthaswamy et al 2012). Many of the DNA-based food authentication assays are multiplexed end-point PCR assays (Dalmasso et al 2004;Zha et al 2010;Ghovvati et al 2009;Rodriguez et al 2003). A majority of these protocols require subsequent steps such as restriction enzyme digestion or gel electrophoresis for species identification after PCR.…”

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confidence: 99%

A nuclear DNA-based species determination and DNA quantification assay for common poultry species

Satkoski

Premasuthan

et al. 2012

J Food Sci Technol

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DNA testing for food authentication and quality control requires sensitive species-specific quantification of nuclear DNA from complex and unknown biological sources. We have developed a multiplex assay based on TaqMan® realtime quantitative PCR (qPCR) for species-specific detection and quantification of chicken (Gallus gallus), duck (Anas platyrhynchos), and turkey (Meleagris gallopavo) nuclear DNA. The multiplex assay is able to accurately detect very low quantities of species-specific DNA from single or multispecies sample mixtures; its minimum effective quantification range is 5 to 50 pg of starting DNA material. In addition to its use in food fraudulence cases, we have validated the assay using simulated forensic sample conditions to demonstrate its utility in forensic investigations. Despite treatment with potent inhibitors such as hematin and humic acid, and degradation of template DNA by DNase, the assay was still able to robustly detect and quantify DNA from each of the three poultry species in mixed samples. The efficient species determination and accurate DNA quantification will help reduce fraudulent food labeling and facilitate downstream DNA analysis for genetic identification and traceability. ). The integration of species determination and nuclear DNA quantification into a single assay will streamline downstream genotyping analysis, and improve the quality of the DNA profiles while conserving reagents (Evans et al. 2007;Lindquist et al. 2011;Kanthaswamy et al. 2012). Many of the DNA-based food authentication assays are multiplexed end-point PCR assays (Dalmasso et al. 2004;Zha et al. 2010;Ghovvati et al. 2009;Rodriguez et al. 2003). A majority of these protocols require subsequent steps such as restriction enzyme digestion or gel electrophoresis for species identification after PCR. Endpoint PCR assays with a quantification component have been developed, but amplicon quantities are typically determined using imaging software analysis of fluorescence intensities of electrophoresis gel bands (Soares et al. 2010).Like end-point PCR, real-time quantitative PCR (qPCR) technology allows multiplexing, and is amenable to detecting multiple DNA targets that allow the simultaneous identification of DNA originating from multiple species. Moreover, the real-time DNA quantification feature of qPCR technology facilitates the efficient and accurate quantification of template DNA. While simplifying data collection and analysis, qPCR's ability to amplify, identify, and quantify in a single step effectively minimizes turnaround time and exposure to contamination and errors ).Electronic supplementary material The online version of this article

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A multiplex PCR assay for fraud identification of deer products

Cited by 33 publications

References 15 publications

Development and validation of fast Real-Time PCR assays for species identification in raw and cooked meat mixtures

Development and validation of fast Real-Time PCR assays for species identification in raw and cooked meat mixtures

Identification of Puffer Fish of the GenusLagocephalus: L. lunaris, L. spadiceusandL. inermis, Using Multiplex PCR

A nuclear DNA-based species determination and DNA quantification assay for common poultry species

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