Practicable Group Testing Method to Evaluate Weight/Weight GMO Content in Maize Grains

Mano, Junichi; Yanaka, Yuka; Ikezu, Yoko; Onishi, Mari; Futo, Satoshi; Minegishi, Yasutaka; Ninomiya, Kenji; Yotsuyanagi, Yuichi; Spiegelhalter, Frank; Akiyama, Hiroshi; Teshima, Reiko; Hino, Akihiro; Naito, Shigehiro; Koiwa, Tomohiro; Takabatake, Reona; Furui, Satoshi; Kitta, Kazumi

doi:10.1021/jf200212v

Cited by 20 publications

(18 citation statements)

References 16 publications

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“…The GMO content is then evaluated statistically based on the qualitative results from multiple small pools of grains ( Fig. 3) (Mano et al 2011). The testing method consists of a sample pretreatment step in which a group of 20 maize kernels is ground in a lysis buffer by a household food processor.…”

Section: Group Testing and Its Validationmentioning

confidence: 99%

Development and Standardization of Analytical Methods for Increasing Varieties of Genetically Modified Crops

Mano

Takabatake

Kitta

2020

JARQ

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The varieties of commercially available genetically modified (GM) crops are rapidly increasing, and this situation demands analytical methods capable of detecting recently developed GM crops. Here we review our research activities to develop and validate new analytical methods for recently distributed GM crops. For the screening analysis of GM content in analytical samples, we developed real-time PCR-based quantitative methods for two GM maize events, MIR604 and MIR162. To accurately analyze GM content irrespective of the commingling of stacked GM events, we developed the group testing method. For the comprehensive analysis of various GM events, the real-time PCR array method was established. In November 2016, the Consumer Affairs Agency of Japan released the standard testing manual including these new testing methods to ensure the validity of the food labeling system in Japan. Given the expected increase in the number of GM events to be analyzed in the future, we need to keep working toward the realization of simple and comprehensive detection and quantification methods that can be used for the increasing number of these events.

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Section: Group Testing and Its Validationmentioning

confidence: 99%

Development and Standardization of Analytical Methods for Increasing Varieties of Genetically Modified Crops

Mano

Takabatake

Kitta

2020

JARQ

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“…In this study, we adopted plasmid DNA theoretically adjusted to 40 copies as the DNA template in the IPC reaction. We used IPC 1-5 ′ , IPC 1-3 ′ , IPC 1-Taq and pArt plasmid as described in our previous report 8) as the forward primer, reverse primer, TaqMan probe and DNA template, respectively. To confirm the absence of apparent PCR inhibition, we set the criterion that the DNA amplification curves of the IPC reaction should cross the threshold line.…”

Section: Applicability Evaluation Of Direct Real-time Pcr Analysismentioning

confidence: 99%

“…For this reason, we investigated a direct real-time PCR analysis based on realtime monitoring of DNA amplification directly from the crude cell lysate without any purification step. We previously reported a simple pretreatment method to prepare crude cell lysates from maize grains for GMO analysis 8) .…”

Section: Introductionmentioning

confidence: 99%

“…The adjusted composition of the reagent makes DNA amplification more resistant to potential PCR inhibitors including sur-* kaz@affrc.go.jp factants used for cell lysis. In the present study, we adopted the previously reported pretreatment procedure 8) and the newly developed master mix reagent as basal techniques for the direct real-time PCR system (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

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Development of Direct Real-Time PCR System Applicable to a Wide Range of Foods and Agricultural Products

Mano

Hatano²,

Futo³

et al. 2014

Journal of the Food Hygienics Society of Japan

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To improve the efficiency of DNA analysis of foods and agricultural products, we investigated a direct real-time PCR based on the real-time monitoring of DNA amplification directly from crude cell lysates of analytical samples. We established a direct real-time PCR system comprising sample pretreatment with a specified lysis buffer and real-time PCR using the developed master mix reagent. No PCR inhibition was observed in the analysis of crude cell lysates from 50 types of samples, indicating that the direct real-time PCR system is applicable to a wide range of materials. The specificity of the direct real-time PCR was evaluated by means of a model assay system for single nucleotide discrimination. Even when crude cell lysates coexisted in the reaction mixtures, the primer selectivity was not affected, suggesting that the sequence specificity of the direct real-time PCR was equivalent to that of PCR from purified DNA templates. We evaluated the sensitivity and quantitative performance of the direct real-time PCR using soybean flour samples including various amounts of genetically modified organisms. The results clearly showed that the direct real-time PCR system provides sensitive detection and precise quantitation.

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“…This detection system has already been implemented in Japan as an official GM maize detection method [19]. Moreover, a GMO content evaluation method based on group testing strategy [20][21][22] was recently developed [23]. In this method, GMO content is statistically evaluated based on qualitative PCR for multiple small portions, consisting of 20 maize kernels.…”

Section: Introductionmentioning

confidence: 99%

A novel trait-specific real-time PCR method enables quantification of genetically modified (GM) maize content in ground grain samples containing stacked GM maize

Akiyama

Nakamura

Sakata

et al. 2014

Eur Food Res Technol

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The GMO contents of two genes were quantified using a plasmid calibrant and summed for quantification of total GMO content. The trait-specific method revealed lower biases for examination of test samples containing stacked GM maize compared with the event-specific method. Our results clearly show that the trait-specific method is not only simple and cost-effective, but also useful in quantifying the GMO content in ground grain samples containing stacked GM maize, which are expected to be major events in the near future. The developed method would be the only feasible way to conduct the quantification of GMO content in the ground maize samples containing stacked GM maize for the verification of the labeling regulation.

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Practicable Group Testing Method to Evaluate Weight/Weight GMO Content in Maize Grains

Cited by 20 publications

References 16 publications

Development and Standardization of Analytical Methods for Increasing Varieties of Genetically Modified Crops

Development and Standardization of Analytical Methods for Increasing Varieties of Genetically Modified Crops

Development of Direct Real-Time PCR System Applicable to a Wide Range of Foods and Agricultural Products

A novel trait-specific real-time PCR method enables quantification of genetically modified (GM) maize content in ground grain samples containing stacked GM maize

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