Development of the Visual Loop-Mediated Isothermal Amplification Assays for Seven Genetically Modified Maize Events and Their Application in Practical Samples Analysis

Chen, Lili; Guo, Jinchao; Wang, Qidi; Kai, Guoyin; Liu, Yang

doi:10.1021/jf200459s

Cited by 57 publications

(31 citation statements)

References 24 publications

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“…Chen developed a LAMP method to detect specific GMOs, based on the event-specific sequences, in seven GM maize events. The detection limit of this method reached four copies of the maize haploid genomic DNA [15]. Positive LAMP results can be easily observed both by gel electrophoresis or by the naked eye through a color change of the reaction mixture with SYBR green I insertion.…”

Section: Journal Of Food and Nutrition Researchmentioning

confidence: 93%

Rapid Detection of Genetically Modified Ingredients in Soybean Products by Real-Time Loop-Mediated Isothermal Amplification

Di¹,

Shi²,

Shen³

et al. 2014

JFNR

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A real-time turbidimeter loop-mediated isothermal amplification (LAMP) assay was developed for the rapid, sensitive, and convenient detection of genetically modified (GM) ingredients in soybean products obtained from local supermarkets in China. Specific primers were designed based on the published sequence of lectin, nos terminator and endogenous genes of two GM soybean strains (GTS40-3-2 and MON89788). The total genomic DNA of soybean-made products was extracted by cetyltrimethylammonium bromide (CTAB) method. Then, the genomic DNA was used as template for LAMP assay. The results showed that this real-time turbidimeter LAMP assay was specific and sensitive enough to amplify DNA from soybean products. The limit of detection (LOD) of lectin and MON89788 was 0.01% (weight of GM soybean/weight of non-GM soybean), roughly equivalent to four copies of soybean genomic DNA in DNA mixture within one hour; and the LOD of nos and GTS40-3-2 was 0.1%, roughly equivalent to forty copies of soybean genomic DNA within 1 hour. The sensitivity reaches to 0.01%, which meets the requirements of the European Union (EU). In practical application, the total detectable ratio in soybeanmade products was the same as that of PCR, suggesting that LAMP method was accurate and sensitive.

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Section: Journal Of Food and Nutrition Researchmentioning

confidence: 93%

Rapid Detection of Genetically Modified Ingredients in Soybean Products by Real-Time Loop-Mediated Isothermal Amplification

Di¹,

Shi²,

Shen³

et al. 2014

JFNR

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“…The identification of seven GM maize events (DAS-59122-7, T25, Bt176, TC1507, MON810, Bt11, and MON863) with LAMP was reported in 2011, and the LOD was as low as four copies of maize haploid genomic DNA, with the exception of MON 810 [27]. In addition, LAMP has been used to detect GM cottons [28], the sensitivity of LAMP was found to be 100-fold higher than that of PCR.…”

Section: Detection Of Gm Eventsmentioning

confidence: 99%

Loop-Mediated Isothermal Amplification (LAMP) Assay for GMO on: Recent Progresses and Future Perspectives

Li¹,

Ji²,

Zhao³

et al. 2015

OALib

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As more and more genetically modified (GM) crops are approved for commercialization and planting, safety issues of GM crops have become hot topics worldwide. For both regulatory and academic purposes, development of rapid, economic and effective on-site detection methods for GM components is indispensible. Up to now, the most effective and sensitive techniques used for GMO detection are based on polymerase chain reaction (PCR). PCR method needs expensive, heavy instruments and gel electrophoresis. Therefore, it is commonly used in laboratory test, and unsuitable for on-site detection. Loop-mediated isothermal amplification (LAMP), an isothermal nucleic acid amplification technique, has been extensively used in many areas such as food safety and clinic diagnosis. Advantageous characteristics of LAMP, such as high specificity and sensitivity, simple operation, low cost, eye visualization, particularly free of special equipment, render it with high potential to be used for GMO on-site detection. In this review, we summarized current status of the application of LAMP in GMO detection, and discussed possible improvements needed for its adaptability regarding to on-site GMO detection. Hopefully, the information present here would facilitate the practical risk assessment of GMO.

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“…Due to the rapid increase of new GM events, screen-specific and event-specific methods are mainly used in routine laboratory analysis, and screen-specific, gene-specific, and construct-specific test are all screening tests. Several modified LAMP assays have been developed and used in GMO detection, including the event-specific visual LAMP assays of two GM soybean events (GTS40-3-2 and MON89788) [12] and seven GM maize events (DAS-59122-7, T25, BT176, TC1507, MON810, BT11, and MON863) [14]. Two screen-specific LAMP assays of CaMV35S promoter and NOS terminator were also developed [15].…”

Section: Introductionmentioning

confidence: 99%

GMO detection in food and feed through screening by visual loop-mediated isothermal amplification assays

Wang

Quan

et al. 2015

Anal Bioanal Chem

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Isothermal DNA/RNA amplification techniques are the primary methodology for developing on-spot rapid nucleic acid amplification assays, and the loop-mediated isothermal amplification (LAMP) technique has been developed and applied in the detection of foodborne pathogens, plant/animal viruses, and genetically modified (GM) food/feed contents. In this study, one set of LAMP assays targeting on eight frequently used universal elements, marker genes, and exogenous target genes, such as CaMV35S promoter, FMV35S promoter, NOS, bar, cry1Ac, CP4 epsps, pat, and NptII, were developed for visual screening of GM contents in plant-derived food samples with high efficiency and accuracy. For these eight LAMP assays, their specificity was evaluated by testing commercial GM plant events and their limits of detection were also determined, which are 10 haploid genome equivalents (HGE) for FMV35S promoter, cry1Ac, and pat assays, as well as five HGE for CaMV35S promoter, bar, NOS terminator, CP4 epsps, and NptII assays. The screening applicability of these LAMP assays was further validated successfully using practical canola, soybean, and maize samples. The results suggested that the established visual LAMP assays are applicable and cost-effective for GM screening in plant-derived food samples.

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Development of the Visual Loop-Mediated Isothermal Amplification Assays for Seven Genetically Modified Maize Events and Their Application in Practical Samples Analysis

Cited by 57 publications

References 24 publications

Rapid Detection of Genetically Modified Ingredients in Soybean Products by Real-Time Loop-Mediated Isothermal Amplification

Rapid Detection of Genetically Modified Ingredients in Soybean Products by Real-Time Loop-Mediated Isothermal Amplification

Loop-Mediated Isothermal Amplification (LAMP) Assay for GMO on: Recent Progresses and Future Perspectives

GMO detection in food and feed through screening by visual loop-mediated isothermal amplification assays

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