Antibiotics, which are commonly used to inhibit the growth of microorganisms, play a major role in the food industry. For example, antibiotics are used in the treatment and prevention of bacterial infections. However, the overuse of antibiotics may lead to serious side effects such as chemical poiso...
Antibiotics, which are commonly used to inhibit the growth of microorganisms, play a major role in the food industry. For example, antibiotics are used in the treatment and prevention of bacterial infections. However, the overuse of antibiotics may lead to serious side effects such as chemical poisoning, allergic reactions, vomiting and diarrhea, etc. Therefore, it is crucial to verify whether residual antibiotics in food exceed the maximum residues limit (MRL). MRL definitions are formulated to monitor excessively high concentrations of antibiotics in food by the Food and Drug Administration (FDA), Food and Agriculture Organization of the United Nations / World Health Organization, (FAO/WHO), or the Korea Food and Drug Administration (KFDA). In order to detect residual antibiotics in food products, a variety of methods such as high-performance liquid chromatography (HPLC), enzyme-linked immunosorbent assay (ELISA), and capillary electrophoresis (CE) have been used. HPLC is the most widely used method, which has high sensitivity and selectivity, but this method needs expensive sample preparation and equipment. ELISA is limited in practice because of its reliance on the surrounding environment and CE also needs expensive instrumentation. As a consequence, the development of inexpensive, rapid, accurate and sensitive methods which can detect residual antibiotics in food are required.
Aptamers are single-stranded DNA (ssDNA) which has been known as a probe for proteins, cells and small molecules, can specifically bind to target molecules for high affinity. Because of their exclusive advantages such as low cost, easy synthesis, and high chemical stability, a various kind of aptamer based sensor has been developed. Also, aptamers for some antibiotics have been developed and used in a sensing method utilizing diverse methods such as electrochemical spectroscopy, colorimetry, and surface plasmon resonance (SPR) spectroscopy.
Graphene oxide (GO) which has many oxygen-containing groups on its surface is widely used in a sensing field because of its unique characteristics, such as facile surface modification, large surface area, and strong photoluminescence. Moreover, because of non-radioactive electronic excitation energy transfer, GO could be used to develop fluorescence resonance energy transfer (FRET) sensor. Fluorescent DNA aptamer can be adsorbed on GO by π-π stacking, resulting in quenching of the fluorescent labeled aptamer. While the addition of antibiotics desorb each paired sequence, the fluorescence intensity is increased. Furthermore, to achieve sensitive and multiplexed detection, the cyclic enzymatic amplification method have been developed using endonuclease.
In this study, ssDNA aptamers targeting to kanamycin, ampicillin, and sulfadimethoxine are labeled with FAM, Cy-3 and Cy-5. After aptamers, which were adsorbed on GO, are quenched by FRET, each antibiotic binds to aptamer resulting in fluorescence with different wavelength. Additionally, this FRET sensor using endonuclease can have signal amplification effect resulting in high sensitivity.
Also, another research about developing aptamers to diagnose subtropical infectious diseases (Dengue virus, Chikungunya virus) and characterization of aptamers was performed.
Antibiotics, which are commonly used to inhibit the growth of microorganisms, play a major role in the food industry. For example, antibiotics are used in the treatment and prevention of bacterial infections. However, the overuse of antibiotics may lead to serious side effects such as chemical poisoning, allergic reactions, vomiting and diarrhea, etc. Therefore, it is crucial to verify whether residual antibiotics in food exceed the maximum residues limit (MRL). MRL definitions are formulated to monitor excessively high concentrations of antibiotics in food by the Food and Drug Administration (FDA), Food and Agriculture Organization of the United Nations / World Health Organization, (FAO/WHO), or the Korea Food and Drug Administration (KFDA). In order to detect residual antibiotics in food products, a variety of methods such as high-performance liquid chromatography (HPLC), enzyme-linked immunosorbent assay (ELISA), and capillary electrophoresis (CE) have been used. HPLC is the most widely used method, which has high sensitivity and selectivity, but this method needs expensive sample preparation and equipment. ELISA is limited in practice because of its reliance on the surrounding environment and CE also needs expensive instrumentation. As a consequence, the development of inexpensive, rapid, accurate and sensitive methods which can detect residual antibiotics in food are required.
Aptamers are single-stranded DNA (ssDNA) which has been known as a probe for proteins, cells and small molecules, can specifically bind to target molecules for high affinity. Because of their exclusive advantages such as low cost, easy synthesis, and high chemical stability, a various kind of aptamer based sensor has been developed. Also, aptamers for some antibiotics have been developed and used in a sensing method utilizing diverse methods such as electrochemical spectroscopy, colorimetry, and surface plasmon resonance (SPR) spectroscopy.
Graphene oxide (GO) which has many oxygen-containing groups on its surface is widely used in a sensing field because of its unique characteristics, such as facile surface modification, large surface area, and strong photoluminescence. Moreover, because of non-radioactive electronic excitation energy transfer, GO could be used to develop fluorescence resonance energy transfer (FRET) sensor. Fluorescent DNA aptamer can be adsorbed on GO by π-π stacking, resulting in quenching of the fluorescent labeled aptamer. While the addition of antibiotics desorb each paired sequence, the fluorescence intensity is increased. Furthermore, to achieve sensitive and multiplexed detection, the cyclic enzymatic amplification method have been developed using endonuclease.
In this study, ssDNA aptamers targeting to kanamycin, ampicillin, and sulfadimethoxine are labeled with FAM, Cy-3 and Cy-5. After aptamers, which were adsorbed on GO, are quenched by FRET, each antibiotic binds to aptamer resulting in fluorescence with different wavelength. Additionally, this FRET sensor using endonuclease can have signal amplification effect resulting in high sensitivity.
Also, another research about developing aptamers to diagnose subtropical infectious diseases (Dengue virus, Chikungunya virus) and characterization of aptamers was performed.
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