Development of nucleic acid coated nanoparticle based lateral flow assay for e.coli detection

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2019
Bingöl, Doğa
Foodborne diseases have been a serious issue for all societies since the beginning of humanity. Thousands of people suffer from foodborne diseases (FBD) every year according to WHO. These diseases are important due to their high frequency and the expenditures that they affect community negatively. Both developing and developed countries can be affected from large outbreaks of FBD. Although most strains of Escherichia coli are beneficial for host microbiota, some serotypes can lead to serious foodborne illnesses in their host. O157:H7 is one of the most virulent strains of E.coli and fast detection of E.coli O17:H7 is a fundamental aim to help raise the food safety and diminish outbreaks globally. Recently, nanoparticle based biosensors have drawn attention of scientists because of their quickness, easiness, sensitivity and specificity. The popularity of system composed of target aiming controlled release of cargo has dramatically increased over t In this study, hybridization was triggered to detect target oligonucleotide designed from eaeA gene which is found in E.coli O17:H7. Single stranded probe oligonucleotide with perfect matching for eaeA target was used to cap 3,3’,5,5’- Tetramethylbenzidine (TMB) loaded silica nanoparticles. As a result of hybridization between probe and target, horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) can oxidize TMB which was liberated. Optimization studies were done in order to obtain proper colorimetric reaction on LFA. Therefore, target concentration could be selectively sensed by probes until it was 7.5 μM.he past few years. In this study, a novel Probe-gated system was assessed for the detection of eaeA target gene. In the presence of target, cargo is delivered selectively from the pores of nanoparticles.