Traditional transgenic detection methods require high test conditions and struggle to be both sensitive and efficient.In this study,a one-tube dual recombinase polymerase amplification(RPA)reaction system for CP4-EPSP...Traditional transgenic detection methods require high test conditions and struggle to be both sensitive and efficient.In this study,a one-tube dual recombinase polymerase amplification(RPA)reaction system for CP4-EPSPS and Cry1Ab/Ac was proposed and combined with a lateral flow immunochromatographic assay,named“Dual-RPA-LFD”,to visualize the dual detection of genetically modified(GM)crops.In which,the herbicide tolerance gene CP4-EPSPS and the insect resistance gene Cry1Ab/Ac were selected as targets taking into account the current status of the most widespread application of insect resistance and herbicide tolerance traits and their stacked traits.Gradient diluted plasmids,transgenic standards,and actual samples were used as templates to conduct sensitivity,specificity,and practicality assays,respectively.The constructed method achieved the visual detection of plasmid at levels as low as 100 copies,demonstrating its high sensitivity.In addition,good applicability to transgenic samples was observed,with no cross-interference between two test lines and no influence from other genes.In conclusion,this strategy achieved the expected purpose of simultaneous detection of the two popular targets in GM crops within 20 min at 37°C in a rapid,equipmentfree field manner,providing a new alternative for rapid screening for transgenic assays in the field.展开更多
Industry progressing caused the pollutants into the surface and groundwater,including organic pollutants such as phenylenediamine(PD)and heavy metals such as Cr(Ⅵ).Herein,a simple"three-in-one"strategy base...Industry progressing caused the pollutants into the surface and groundwater,including organic pollutants such as phenylenediamine(PD)and heavy metals such as Cr(Ⅵ).Herein,a simple"three-in-one"strategy based on MOF-199 was demonstrated.The MOF-199 catalytic system could be used for the rapid detection of Cr(Ⅵ)and o-phenylenediamine(OPD)in linear ranges of 0.5-50.0μmol/L and 7.0-250.0μmol/L,with the limits of detection of 0.1 and 0.5μmol/L,respectively.This method can also efficiently discriminate PD isomers in just 15 min.Meanwhile,Cr(Ⅵ)was reduced to Cr(Ⅲ)and the elimination of OPD was carried out.Given simple synthesis,high sensitivity,easy operation,and low cost,this method presented a potential platform for on-site detection of pollutants in water samples.展开更多
基金supported by the Scientific and Innovative Action Plan of Shanghai(21N31900800)Shanghai Rising-Star Program(23QB1403500)+4 种基金the Shanghai Sailing Program(20YF1443000)Shanghai Science and Technology Commission,the Belt and Road Project(20310750500)Talent Project of SAAS(2023-2025)Runup Plan of SAAS(ZP22211)the SAAS Program for Excellent Research Team(2022(B-16))。
文摘Traditional transgenic detection methods require high test conditions and struggle to be both sensitive and efficient.In this study,a one-tube dual recombinase polymerase amplification(RPA)reaction system for CP4-EPSPS and Cry1Ab/Ac was proposed and combined with a lateral flow immunochromatographic assay,named“Dual-RPA-LFD”,to visualize the dual detection of genetically modified(GM)crops.In which,the herbicide tolerance gene CP4-EPSPS and the insect resistance gene Cry1Ab/Ac were selected as targets taking into account the current status of the most widespread application of insect resistance and herbicide tolerance traits and their stacked traits.Gradient diluted plasmids,transgenic standards,and actual samples were used as templates to conduct sensitivity,specificity,and practicality assays,respectively.The constructed method achieved the visual detection of plasmid at levels as low as 100 copies,demonstrating its high sensitivity.In addition,good applicability to transgenic samples was observed,with no cross-interference between two test lines and no influence from other genes.In conclusion,this strategy achieved the expected purpose of simultaneous detection of the two popular targets in GM crops within 20 min at 37°C in a rapid,equipmentfree field manner,providing a new alternative for rapid screening for transgenic assays in the field.
基金supported by the National Natural Science Foundation of China(NSFC)Fund(22174058,U21A20282)Sci-tech plan projects of Gansu province(22CX3GA023).
文摘Industry progressing caused the pollutants into the surface and groundwater,including organic pollutants such as phenylenediamine(PD)and heavy metals such as Cr(Ⅵ).Herein,a simple"three-in-one"strategy based on MOF-199 was demonstrated.The MOF-199 catalytic system could be used for the rapid detection of Cr(Ⅵ)and o-phenylenediamine(OPD)in linear ranges of 0.5-50.0μmol/L and 7.0-250.0μmol/L,with the limits of detection of 0.1 and 0.5μmol/L,respectively.This method can also efficiently discriminate PD isomers in just 15 min.Meanwhile,Cr(Ⅵ)was reduced to Cr(Ⅲ)and the elimination of OPD was carried out.Given simple synthesis,high sensitivity,easy operation,and low cost,this method presented a potential platform for on-site detection of pollutants in water samples.