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A powerless on-the-spot detection protocol for transgenic crops within 30 min, from leaf sampling up to results

文献类型： 外文期刊

作者： Wang, Liu 1 ; Wang, Rui 1 ; Yu, Yonghua 1 ; Zhang, Fang 1 ; Wang, Xiaofu 2 ; Ying, Yibin 1 ; Wu, Jian 1 ; Xu, Junfeng 2 ;

作者机构： 1.Zhejiang Univ, Coll Biosyst Engn & Food Sci, Hangzhou 310058, Zhejiang, Peoples R China

2.Zhejiang Acad Agr Sci, Hangzhou 310021, Zhejiang, Peoples R China

关键词： Crude DNA extract;Unpowered device;Loop-mediated isothermal amplification;Visual detection;Transgenic crop detection

期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY （ 影响因子：4.142； 五年影响因子：3.863 ）

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收录情况： SCI

摘要： The requirement of power-dependent instruments or excessive operation time usually restricts current nucleic acid amplification methods from being used for detection of transgenic crops in the field. In this paper, an easy and rapid detection method which requires no electricity supply has been developed. The time-consuming process of nucleic acid purification is omitted in this method. DNA solution obtained from leaves with 0.5 M sodium hydroxide (NaOH) can be used for loop-mediated isothermal amplification (LAMP) only after simple dilution. Traditional instruments like a polymerase chain reaction (PCR) amplifier and water bath used for DNA amplification are abandoned. Three kinds of dewar flasks were tested and it turned out that the common dewar flask was the best. Combined with visual detection of LAMP amplicons by phosphate (Pi)-induced coloration reaction, the whole process of detection of transgenic crops via genetically pure material (leaf material of one plant) could be accomplished within 30 min. The feasibility of this method was also verified by analysis of practical samples.

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