Study And Application Of Fluorescent Biosensor For The Detection Of Carbamate Pesticide Residues

Recent years,the issue of pesticide residues in food have caused more attention all over the world.The toxicosis incidents of human and animal resulted from taking food which contains high level of pesticide residues are happened usually,which are harming to food security. The traditional methods of detection of pesticide residues such as GC,HPLC,GC-MS and LC-MS,analysis instrument is expensive, sample preparation is complicated, testing cycle lasts long,difficult to solve the problem of pesticide residues effectively.Therefore,research and development a convenient,rapid,sensitive and practical technique such as fluorescent biosensor technique to adapt to the produce and sale status of agriculture product for inspection pesticide residues are urgent demands in our country today.At present, detection of pesticide residues by bioluminescence-based biosensor is a hot spot.In this dissertation,we identified the function of two carbamate degrading bacteria,and successfully constructed the metagenomic library,where we found out407bp DNA fragment which is able to identify carbamate pesticides.Thus.we constructed H12-E cell-free system fluorescent biosensor and standard methods for the detection of carbamate pesticide residues, the limit of detection (LOD) was1fg/L.The standard curve equation for carbofuran is y=1.0315Ln(x)+0.8788,R2=0.991,for carbaryl is y=0.9685Ln(x)+1.554.R2=0.9925,the recovery rate is86.7%-102.4%.Some detections of carbamate residues in vegetables have been done via H12-E fluorescent biosensor.The result reveals H12-E fluorescent biosensor can detect the total amount of carbamate pesticide residues rapidly and accurately,so the pollution which is caused by a variety of pesticide residue can be solved effectively.In summary,H12-E fluorescent biosensor has wide application prospect and great value for development.