Quantification of DNA fragmentation in processed foods using real-time PCR

Mano, Junichi; Nishitsuji, Yasuyuki; Kikuchi, Yoshihiro; Fukudome, Shin-ichi; Hayashida, Takuya; Kawakami, Hiroyuki; Kurimoto, Youichi; Niimi, Akio; Kondo, Kazunari; Teshima, Reiko; Takabatake, Reona; Kitta, Kazumi

doi:10.1016/j.foodchem.2017.01.064

Cited by 41 publications

(36 citation statements)

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“…Indeed, previous studies showed that real‐time PCR is 10‐ to 100‐fold more sensitive than conventional PCR . Thus, real‐time PCR is suitable for analysis of food that has undergone intensive processing and contains very small amounts of DNA . We carried out standard curve analyses to quantify real‐time PCR assays using the SCAR markers.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, DNA fragmentation (to sizes less than 200 bp) frequently occurs in processed food such as pressed oils, cooked foods and ground material, making it difficult to detect DNA . DNA fragmentation caused by physical force, heat, pH change or enzymatic activity decreases amplification efficiency because it is difficult for primers to bind highly fragmented DNA in the annealing step of PCR . Consistent with this, some quantitative assays yield different results from raw materials and processed foods .…”

Section: Introductionmentioning

confidence: 99%

“…DNA fragmentation caused by physical force, heat, pH change or enzymatic activity decreases amplification efficiency because it is difficult for primers to bind highly fragmented DNA in the annealing step of PCR . Consistent with this, some quantitative assays yield different results from raw materials and processed foods . Therefore, accurate species discrimination is necessary to confirm the quality of processed food, and to this end it is necessary to develop analytic methods with high sensitivity.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, accurate species discrimination is necessary to confirm the quality of processed food, and to this end it is necessary to develop analytic methods with high sensitivity. A recent study reported that the degree of DNA fragmentation affects the amplification efficiency of real‐time PCR . Real‐time PCR assays have a very low limit of detection (LOD), and it is possible to discriminate species even from fragmented DNA .…”

Section: Introductionmentioning

confidence: 99%

“…A recent study reported that the degree of DNA fragmentation affects the amplification efficiency of real‐time PCR . Real‐time PCR assays have a very low limit of detection (LOD), and it is possible to discriminate species even from fragmented DNA . In addition, SYBR Green‐based real‐time PCR assays are less costly than probe‐based assays …”

Section: Introductionmentioning

confidence: 99%

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Development of conventional PCR and real‐time PCR assays to discriminate the origins of Chinese pepper oil and herbal materials from Zanthoxylum

et al. 2018

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BACKGROUND To ensure the safety, quality and therapeutic efficacy of processed foods and herbal medicines, it is important to identify and discriminate economically motivated adulterants. Zanthoxylum schinifolium is sold at a higher price than other Zanthoxylum species and is frequently adulterated with closely related Zanthoxylum species because of its high demand as a Korean food ingredient and medicinal material in markets. In addition, the pericarps of three Zanthoxylum species ( Z. schinifolium , Z. bungeanum and Z. piperitum ) are defined as herbal medicine Zanthoxyli Pericarpium in Korean pharmacopoeias, but not Z. piperitum in Chinese pharmacopoeias. Further confusion arises in the morphological similarity between Z. armatum (adulterant) and Z. bungeanum . Therefore, the aim of this study was to develop a sequence characterized amplified region (SCAR) marker for discrimination of four Zanthoxylum species. RESULTS With the goal of developing rapid and reliable tools for genetic discrimination of authentic Zanthoxyli Pericarpium, we designed species‐specific SCAR markers, based on ITS2 sequences, that generate amplicons of less than 200 bp. Using these markers, we established both conventional and real‐time PCR assay methods capable of differentiating samples at the species level. We validated the ability of SCAR markers to authenticate edible oil and herbal medicine, and confirmed that some herbal medicines contaminated with Z. armatum are being distributed as Zanthoxyli Pericarpium in Korean and Chinese markets. CONCLUSIONS The SCAR markers and PCR methods described represent powerful tools for protecting against adulteration and ensuring standardization of processed foods and herbal medicine. © 2018 The Authors. Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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