title: One simple DNA extraction device and its combination with modified visual loop-mediated isothermal amplification for rapid on-field detection of genetically modified organisms.
abstract: Quickness, simplicity, and effectiveness are the three major criteria for establishing a good molecular diagnosis method in many fields. Herein we report a novel detection system for genetically modified organisms (GMOs), which can be utilized to perform both on-field quick screening and routine laboratory diagnosis. In this system, a newly designed inexpensive DNA extraction device was used in combination with a modified visual loop-mediated isothermal amplification (vLAMP) assay. The main parts of the DNA extraction device included a silica gel membrane filtration column and a modified syringe. The DNA extraction device could be easily operated without using other laboratory instruments, making it applicable to an on-field GMO test. High-quality genomic DNA (gDNA) suitable for polymerase chain reaction (PCR) and isothermal amplification could be quickly isolated from plant tissues using this device within 15 min. In the modified vLAMP assay, a microcrystalline wax encapsulated detection bead containing SYBR green fluorescent dye was introduced to avoid dye inhibition and cross-contaminations from post-LAMP operation. The system was successfully applied and validated in screening and identification of GM rice, soybean, and maize samples collected from both field testing and the Grain Inspection, Packers, and Stockyards Administration (GIPSA) proficiency test program, which demonstrated that it was well-adapted to both on-field testing and/or routine laboratory analysis of GMOs.
authors.affiliation: National Center for Molecular Characterization of Genetically Modified Organisms, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
authors.forename: Miao
authors.initials: M
authors.lastname: Zhang
authors.forename: Yinan
authors.initials: Y
authors.lastname: Liu
authors.forename: Lili
authors.initials: L
authors.lastname: Chen
authors.forename: Sheng
authors.initials: S
authors.lastname: Quan
authors.forename: Shimeng
authors.initials: S
authors.lastname: Jiang
authors.forename: Dabing
authors.initials: D
authors.lastname: Zhang
authors.forename: Litao
authors.initials: L
authors.lastname: Yang
mesh_terms: D052160:Benzothiazoles; D017931:DNA Primers / Q000737:chemistry; D018744:DNA, Plant / Q000032:analysis* / Q000302:isolation & purification; D003959:Diamines; D021141:Nucleic Acid Amplification Techniques; D009930:Organic Chemicals / Q000737:chemistry; D018515:Plant Leaves / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011804:Quinolines; D012639:Seeds / Q000235:genetics
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D018744:DNA, Plant; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I