A peach (Prunus persica)-specific gene, Lhcb2, used as an endogenous reference gene for qualitative and real-time quantitative PCR to detect fruit products

Shang, Ying; Zhu, Pengyu; Huang, Kunlun; Liu, Weihong; Tian, Wenying; Luo, Yunbo; Xu, Wentao

doi:10.1016/j.lwt.2013.08.009

Cited by 20 publications

(13 citation statements)

References 29 publications

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“…All qualitative LAMP amplicons of identical size were obtained for all tested mammalian varieties, which demonstrated that there were no major sequence differences between the 15 mammals. The recovered products should be sequenced to make sure the conservative 20 , and the sequences of the fifteen products were identical. The qualitative results were consistent with the results that we obtained from the quantitative LAMP approach, exhibiting fluorescent signals only in the mammalian samples and not the other tested species ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Low copy genes generally have less variation; therefore, a single-copy gene is often the best choice for endogenous reference genes 19 . Qualitative and quantitative approaches using endogenous reference genes can be employed to evaluate the quality of the DNA sample, analyze the food source to determine food allergens and determine the ingredients in complex food admixture 20 . Numerous studies have reported the identification of mammalian species in various animal taxa using DNA-based methods.…”

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confidence: 99%

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A smart sealed nucleic acid biosensor based on endogenous reference gene detection to screen and identify mammals on site

Xiang

Wang

et al. 2017

Sci Rep

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The identification of meat adulteration is a hotspot for food research worldwide. In this paper, a smart and sealed biosensor that combines loop-mediated isothermal amplification (LAMP) with a lateral flow device (LFD) was developed, resulting in the universal mammalian assessment on site. First, the highly specific chromosomal Glucagon gene (Gcg) was chosen as the endogenous reference gene, and the LAMP approach provided double-labeled duplex DNA products using FITC- and BIO- modified primers. Then, an LFD strategy was used for specific signal recognition through an immunoassay. Meanwhile, LFD-LAMP was compared to LAMP and real-time LAMP, the results showed consistent high specificity and sensitivity but in a more convenient and easy-to-use system. In addition, the detection limit was as low as 10 pg, which was equivalent to 3~5 copies in mammals. All of the reactions were performed in a sealed system regardless of the amplification process or products recognized. Therefore, the smart design demonstrated significantly high specificity and the ability to detect trace amounts of DNA in complex and processed foods with mammalian meat. As a universal and specific platform for the detection of mammalian DNA, this smart biosensor is an excellent prospect for species identification and meat adulteration.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

A smart sealed nucleic acid biosensor based on endogenous reference gene detection to screen and identify mammals on site

Xiang

Wang

et al. 2017

Sci Rep

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“…Compared with conventional PCR, quantitative PCR technology has realized the leap of PCR from qualitative to quantitative, and it has higher specificity, effective resolution and higher degree of automation, and widely used in many fields such as gene expression research, transgenic research, drug efficacy assessment, pathogen detection and food composition analysis, especially. Therefore, real-time quantitative PCR detection is considered to be an easy-to-use, accurate, specific, sensitive, and quantitative method [13,14,15]. Furthermore, the species-specific diagnostic marker (also denoted as endogenous reference gene in some literatures) is a significant parameter during PCR amplification, which can evaluate the quality of the extracted DNA and provide the means to quantify the amount of the tested DNA substance in the processed food samples [15].…”

Section: Introductionmentioning

confidence: 99%

Identification of the Pol Gene as a Species-Specific Diagnostic Marker for Qualitative and Quantitative PCR Detection of Tricholoma matsutake

Shan

Wang

et al. 2019

Molecules

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Tricholoma matsutake is a rare, precious, and wild edible fungus that could not be cultivated artificially until now. This situation has given way to the introduction of fake T. matsutake commodities to the mushroom market. Among the methods used to detect food adulteration, amplification of species-specific diagnostic marker is particularly important and accurate. In this study, the Pol gene is reported as a species-specific diagnostic marker to identify three T. matsutake varieties and 10 other types of edible mushrooms through qualitative and quantitative PCR. The PCR results did not reveal variations in the amplified region, and the detection limits of qualitative and quantitative PCR were found to be 8 ng and 32 pg, respectively. Southern blot showed that the Pol gene exists as a single copy in the T. matsutake genome. The method that produced the purest DNA of T. matsutake in this study was also determined, and the high-concentration salt precipitation method was confirmed to be the most suitable among the methods tested. The assay proposed in this work is applicable not only to the detection of raw materials but also to the examination of processed products containing T. matsutake.

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“…Modern molecular methods have been widely applied in food authenticity identification (Fajardo, González, Rojas, García, & Martín, ; Marmiroli, Peano, & Maestri, ; Mayer, ; Rodríguez et al., ; Shang et al., ). For instance, DNA is suitable for the detection and analysis of ingredients of processed products because it carries the genetic materials of life and exhibits high stability (Nielsen, ).…”

Section: Introductionmentioning

confidence: 99%

Identification of pyrG Used as an Endogenous Reference Gene in Qualitative and Real‐Time Quantitative PCR Detection of Pleurotus ostreatus

Shi

Shan

Zhuang

et al. 2018

Journal of Food Science

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A peach (Prunus persica)-specific gene, Lhcb2, used as an endogenous reference gene for qualitative and real-time quantitative PCR to detect fruit products

Cited by 20 publications

References 29 publications

A smart sealed nucleic acid biosensor based on endogenous reference gene detection to screen and identify mammals on site

A smart sealed nucleic acid biosensor based on endogenous reference gene detection to screen and identify mammals on site

Identification of the Pol Gene as a Species-Specific Diagnostic Marker for Qualitative and Quantitative PCR Detection of Tricholoma matsutake

Identification of pyrG Used as an Endogenous Reference Gene in Qualitative and Real‐Time Quantitative PCR Detection of Pleurotus ostreatus

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