Transgenic rice Bt63 detection kit and detection method thereof

Abstract

The invention discloses a transgenic rice Bt63 detection kit and a detection method thereof, relating to transgenic rice. The kit comprises a loop mediated isothermal amplification reaction liquid A, a loop mediated isothermal amplification reaction liquid B, a Bst DNA (Deoxyribose Nucleic Acid) polymerase C and a color developing agent D. The method comprises the following steps of: extracting template DNA; performing loop mediated isothermal amplification; and performing color developing detection. Four specially-designed primers and the Bst DNA polymerase with strand displacement activity are used for specially, efficiently and rapidly amplifying DNA under a constant-temperature condition, so that 109 target sequence coping can be realized within one hour. The amplification effect can be observed through a fluorescent dye. A rapid loop mediated isothermal amplification method with high specificity and low cost is used for detecting Bt63 transgenic rice, and can be used for replacing a CRL (Chemical Research Laboratory) method confirmed by the Joint Research Center, and novel scientific bases are provided for food safety supervision.

Description

technical field [0001] The invention relates to transgenic rice, in particular to a transgenic rice Bt63 detection kit and a detection method thereof. Background technique [0002] Bt63 genetically modified rice is rice containing insect-resistant genes, which can resist diseases and increase rice production, but children's food made from this rice may affect the health of infants. From April 15, 2008, the EU requires all rice products imported from China to be inspected by EU-accredited laboratories, and to attach a health certificate confirming that they do not contain "Bt63" (http: / / eur-lex.europa.eu / Lex UriServ / LexUriServ.do?uri=OJ:L:2008:096:0029:0034:EN:PDF). In the past, the detection of Bt63 transgenic rice mainly used ordinary PCR method and fluorescent quantitative PCR method (Xu Ying. The safety of transgenic rice and the progress of nucleic acid detection technology. Anhui Agricultural Sciences, 2009, 37(33): 16240-16242; Wu Xiaohuai, Lu Yong .Using real-time ...

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Problems solved by technology

In the past, the detection of Bt63 transgenic rice mainly used ordinary PCR method and fluorescent quantitative PCR method (Xu Ying. The safety of transgenic rice and the progress of nucleic acid detection technology. Anhui Agricultural Sciences, 2009, 37(33): 16240-16242; Wu Xiaohuai, Lu Yong .Using real-time fluorescent PCR method to detect transgenic rice with Bt [J] Modern Food Science and Technology, 2009 (25): 211-216.), ordinary PCR method needs relatively expensive PCR instrument and gel imaging system, fluorescent quantitative PCR method needs expensive Fluorescent quantitative PCR instrument, grass-roots testing institutions and the majority of food processing enterprises are difficult to popularize the detection method

There is no report on the detection of Bt63 transgenic rice by loop-mediated isothermal amplification

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