title: Rapid visual detection of phytase gene in genetically modified maize using loop-mediated isothermal amplification method.
abstract: Transgenic maize plant expressing high phytase activity has been reported and approved by Chinese government in 2009. Here, we report a highly specific loop-mediated isothermal amplification (LAMP) method to detect the phytase gene in the GMO maize. The LAMP reaction takes less than 20min and the amplification is visible without gel electrophoresis. The detection sensitivity of the LAMP method is about 30 copies of phytase genomic DNA, which is 33.3 times greater than the conventional PCR method with gel electrophoresis. The quantitative detection results showed that the LAMP method has a good linear correlation between the DNA copy number and the associated Tt values over a large dynamic range of template concentration from 6×10(1) to 6×10(7) copies, with a quantification limit of 60 copies. Therefore, the LAMP method is visual, faster, and more sensitive, and does not need special equipment compared to traditional PCR technique, which is very useful for field tests and fast screening of GMO feeds.
authors.affiliation: Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, People's Republic of China. Electronic address: huangxin03@tsinghua.org.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.affiliation: Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, People's Republic of China.
authors.forename: Lili
authors.initials: L
authors.lastname: Chen
authors.affiliation: Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, People's Republic of China.
authors.forename: Jiangmin
authors.initials: J
authors.lastname: Xu
authors.affiliation: Department of Chemistry, Drexel University, Philadelphia, PA 19010, USA.
authors.forename: Hai-Feng
authors.initials: HF
authors.lastname: Ji
authors.affiliation: Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, People's Republic of China.
authors.forename: Shuifang
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, People's Republic of China.
authors.forename: Hongjun
authors.initials: H
authors.lastname: Chen
mesh_terms: D010832:6-Phytase / Q000737:chemistry*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D030781:Organisms, Genetically Modified / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D003313:Zea mays / Q000235:genetics*
chemical_list: D010832:6-Phytase
keywords: DNA extraction; Genetically modified organisms; Loop-mediated isothermal amplification detection; Phytase gene; Rapid visual detection