A Real-time Turbidimeter-based Loopmediated Isothermal Amplification Assay for Rapid Detection of Genetically Modified Maize MON810
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Abstract:
LAMP real-time turbidity method is based on the loop-mediated isothermal amplification, detecting a white precipitate generated during the reaction by a real-time turbidimeter, in order to monitor the whole process of amplification. This technique overcomes the defects of LAMP chromomeric method which could only monitor the end of reaction, therefore becomes more scientific for primers screening and the reaction optimization. Six specific primers were designed for the junction sequences of exogenous gene CryIA(b) and endogenous gene of genetically modified maize MON810. Detection conducted by real-time turbidity method at 63 ℃, the detection sensitivity, specificity and stability were verified. The results indicated that the lowest detection limit of this method was 0.5%, and the coincidence rate compared with the LAMP chromomeric method and the real-time PCR method was 100%, The method had high specificity, stability, accuracy and convenience which was suitable for the rapid detection of genetically modified maize MON810 and has good application value.
