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Colorimetric Detection of Norovirus in Oyster Samples through DNAzyme as a Signaling Probe
Batule, Bhagwan S.,Kim, Seong U,Mun, Hyoyoung,Choi, Changsun,Shim, Won-Bo,Kim, Min-Gon American Chemical Society 2018 Journal of agricultural and food chemistry Vol.66 No.11
<P>Worldwide, norovirus is one of the most associated causes of acute gastroenteritis, which leads to nearly 50 000 child deaths every year in developing countries. Therefore, there is great demand to develop a rapid, low-cost, and accurate detection assay for the foodborne norovirus infection to reduce mortality caused by norovirus. Considering the importance of norovirus, we have demonstrated a highly sensitive and specific colorimetric detection method for analysis of human norovirus genogroups I and II (HuNoV GI and GII) in oyster samples. This is the first report to employ colorimetric HRPzyme-integrated polymerase chain reaction (PCR) for direct norovirus detection from the real shellfish samples. We found that the HRPzyme-integrated PCR method is more sensitive than the gel electrophoresis approach and could detect the HuNoV GI and GII genome up to 1 copy/mL. The specificity of the proposed method was successfully demonstrated for HuNoV GI and GII. Further, we performed testing HuNoVs in the spiked oyster samples, and the HRPzyme-integrated PCR method proved to be an ultrasensitive and selective method for detecting HuNoVs in the real samples. By integration of the proposed method with the portable PCR machine, it would be more reliable to improve food safety by detecting HuNoVs in the different types of shellfish, such as oyster and mussel, at the production field.</P> [FIG OMISSION]</BR>
Ultrafast sonochemical synthesis of protein-inorganic nanoflowers
Batule, Bhagwan S,Park, Ki Soo,Kim, Moon Il,Park, Hyun Gyu Dove Medical Press 2015 INTERNATIONAL JOURNAL OF NANOMEDICINE Vol.10 No.specal
<P>We developed a simple but efficient method to synthesize protein-inorganic hybrid nanostructures with a flower-like shape (nanoflowers), which relies on sonication to facilitate the synthesis of the nanoflowers. With this technique, we synthesized nanoflowers containing laccase as a model protein and copper phosphate within 5 minutes at room temperature. The resulting laccase nanoflowers yielded greatly enhanced activity, stability, and reusability, and their usefulness was successfully demonstrated by applying them in the colorimetric detection of epinephrine. The strategy developed could be used to rapidly synthesize nanoflowers for various applications in biosensor and enzyme catalysis and would expand the utilization of nanoflowers in diverse fields of biotechnology.</P>
Batule, Bhagwan S.,Park, Ki Soo,Gautam, Shreedhar,Cheon, Hong Jae,Kim, Moon Il,Park, Hyun Gyu Elsevier 2019 Sensors and actuators. B Chemical Vol.283 No.-
<P><B>Abstract</B></P> <P>We have found that protein-inorganic hybrid nanoflowers, prepared by an ultrafast sonication-mediated self-assembly of proteins and copper ions, exhibit an intrinsic peroxidase-mimicking activity, which is significantly higher than that of control materials formed in the absence of proteins. By employing glucose oxidase (GOx) as a protein component, the novel synthetic method was applied to construct GOx copper nanoflowers capable of promoting glucose-induced cascade enzymatic reactions. In the presence of target glucose, GOx, entrapped in the hybrid nanoflowers, generates H<SUB>2</SUB>O<SUB>2</SUB> through its catalytic action; this subsequently induces peroxidase-mediated oxidation by the hybrid nanoflowers to convert the selected substrate, Amplex UltraRed (AUR), to a highly fluorescent product. Using this strategy, the target glucose was reliably determined down to 3.5 μM with high selectivity. The practical diagnostic utility of the assay system was also verified by using it to detect glucose in human blood serum. This sonochemical strategy has great potential to be extended for the construction of various oxidative enzyme-inorganic hybrid nanoflowers that are capable of detecting clinically important target molecules.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Sonochemically synthesized protein-inorganic hybrid nanoflowers exhibited an intrinsic peroxidase-like activity. </LI> <LI> Glucose oxidase-copper hybrid nanoflowers possessed dual enzymatic activity. </LI> <LI> The nanoflower-based biosensor showed high selectivity and sensitivity for glucose detection. </LI> <LI> Diverse glucose levels in human blood were successfully determined with excellent reproducibility and reliability. </LI> </UL> </P>