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

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.


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Çam, Dilek; Öktem, Hüseyin Avni (Springer Science and Business Media LLC, 2018-10-25)
Salmonella is among the very important pathogens threating the human and animal health. Rapid and easy detection of these pathogens is crucial. In this context, antibody (Ab) based lateral flow assays (LFAs) which are simple immunochromatographic point of care test kits were developed by gold nanoparticles (GNPs) as labelling agent for Salmonella detection. For that purpose some critical parameters such as reagent concentrations on the capture zones, conjugate concentrations and ideal membrane type needed f...
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Bulut, Onur; Öktem, Hüseyin Avni; Department of Biology (2014)
Foodborne diseases have been a crucial problem for the public health. Various agents transmitted by food cause these diseases. However, Salmonella accounts for most of the cases leading to most of the hospitalization and even death. Therefore, rapid detection of Salmonella is a considerable step in order to improve food safety and minimize outbreaks. Nucleic acid based biosensors are fast, simple, economic, easy-to-use and do not require trained personnel or high-cost equipments when compared to the current...
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Bayraç, Ceren; Öktem, Hüseyin Avni; Eyidoğan, Füsun İnci; Department of Biotechnology (2014)
Food poisoning became one of the most important diseases that threaten to human health. Among all the food-borne pathogens, Salmonella is the most common cause of food-borne infectious disease in the world. Among over 2500 serovars, Salmonella enterica serovar Enteritidis is a common foodborne pathogen associated with human diseases. Limited and time consuming diagnostic techniques such as culturing method or polymerase chain reaction lead to increase in the demands of new detection methods. Therefore, in t...
Recent advances in the biosynthesis of penicillins, cephalosporins and clavams and its regulation
Özcengiz, Gülay (Elsevier BV, 2013-03-01)
The beta-lactam antibiotics have been serving mankind for over 70 years. Despite this old age, they continue to provide health to the world population by virtue of industrial production and discoveries of new secondary metabolite molecules with useful activities. Sales of these remarkable compounds have reached over $20 billion dollars per year. They include penicillins, cephalosporins, cefoxitin, monobactams, clavulanic acid and carbapenems. Strain improvement of the penicillin-producing species of Penicil...
Development of immunoaffinity based detection platforms for food pathogens
Çam, Dilek; Öktem, Hüseyin Avni; Department of Biology (2016)
Salmonella and E.coli, food pathogens, are among the very important pathogens threating the health. Rapid and easy detection of these pathogens is crucial. In this context, lateral flow assay (LFA) platform prepared by gold nanoparticles (GNPs) based on antibody for Salmonella and E.coli O157:H7 was developed in the first part of this study. In second part, single chain variable fragments (scFv) and SNAP-Tagged full IgG (fusion protein) of se155-4 antibody specific to Salmonella were genetically produced. T...
Citation Formats
D. Bingöl, “Development of nucleic acid coated nanoparticle based lateral flow assay for e.coli detection,” Thesis (M.S.) -- Graduate School of Natural and Applied Sciences. Biotechnology., Middle East Technical University, 2019.