Development and evaluation of a novel loop mediated isothermal amplification coupled with TaqMan probe assay for detection of genetically modified organism with NOS terminator

•A novel detection method was developed, coupling LAMP primers with one TaqMan probe.•LOD of LAMP-TaqMan method was 5 copies of targeted DNA per reaction.•This novel detection method was finished in about 20 min. In this study, we aim to develop a novel loop mediated isothermal amplification (LAMP)...

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Veröffentlicht in:Food chemistry 2021-09, Vol.356, p.129684-129684, Article 129684
Hauptverfasser: Yu, Yanbo, Li, Rui, Ma, Zonghua, Han, Meihong, Zhang, Sen, Zhang, Minghui, Qiu, Youwen
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Sprache:eng
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Zusammenfassung:•A novel detection method was developed, coupling LAMP primers with one TaqMan probe.•LOD of LAMP-TaqMan method was 5 copies of targeted DNA per reaction.•This novel detection method was finished in about 20 min. In this study, we aim to develop a novel loop mediated isothermal amplification (LAMP) coupled with TaqMan (LAMP-TaqMan) method for quick qualitative detection of genetically modified organism (GMOs). We designed four LAMP primers and one TaqMan probe for the LAMP-TaqMan detection method to detect the nopaline synthase gene (NOS) terminator in GMOs. This assay enabled the amplification of DNA within ~20 min at a constant temperature of 65 °C. This assay detected as few as five copies of target sequences, which had a high specificity similar to the TaqMan qPCR method. Furthermore, the LAMP-TaqMan detection method was successfully used to amplify and detect DNA from food samples of the major crops (soybean, maize, rice, etc.). In summary, a novel LAMP-TaqMan assay has been developed, which has the similar sensitivity but takes less time than the TaqMan qPCR method. This method offers a novel approach for rapid detection of GMOs in foods.
ISSN:0308-8146
1873-7072
DOI:10.1016/j.foodchem.2021.129684