• Proceedings of the National Institute of Ecology of the Republic of Korea
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• v.1 no.1
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• pp.83-89
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• 2020

Methods for detecting the presence of genetically modified (GM) crops are evolving to comply with legislation and to enhance monitoring by biotechnology companies and regulators. In order to cover a broad range of detection methods for a new GM crop, conventional multiplex PCR methods are required. Based on the genetic information on three GM alfalfa varieties (J101, J163, and KK179), which were recently approved in South Korea, we developed a fast, reliable, and highly specific multiplex polymerase chain reaction (PCR) method with basic PCR equipment and inexpensive reagents. To validate and verify the newly developed multiplex PCR method, we applied a limit of detection assay and random reference material analysis. We also monitored the unintentional environmental release of GM alfalfa in South Korea by performing the multiplex PCR analysis with 91 feral alfalfa specimens collected from 2000 to 2018. Our methodology is a sensitive, simple, quick, and inexpensive tool for detecting and identifying three GM alfalfa varieties.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.681-688
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• 2024

The accurate and rapid detection of methicillin-resistance of Staphylococcus aureus (SA) holds significant clinical importance. However, the methicillin-resistance detection strategies commonly require complicated cell lysis and gene extraction. Herein, we devised a novel colorimetric approach for the sensitive and accurate identification of methicillin-resistance of SA by combining allosteric probe-based target recognition with self-primer elongation-based target recycling. The PBP2a aptamer in the allosteric probe successfully identified the target MRSA, leading to the initiation of self-primer elongation based-cascade signal amplification. The peroxidase-like hemin/G-quadruplex undergo an isothermal autonomous process that effectively catalyzes the oxidation of ABTS2- and produces a distinct blue color, enabling the visual identification of MRSA at low concentrations. The method offers a shorter duration for bacteria cultivation compared to traditional susceptibility testing methods, as well as simplified manual procedures for gene analysis. The overall amplification time for this test is 60 min, and it has a detection limit of 3 CFU/ml. In addition, the approach has exceptional selectivity and reproducibility, demonstrating commendable performance when tested with real samples. Due to its advantages, this colorimetric assay exhibits considerable potential for integration into a sensor kit, thereby offering a viable and convenient alternative for the prompt and on-site detection of MRSA in patients with skin and soft tissue infections.

• Journal of Microbiology and Biotechnology
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• v.27 no.4
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• pp.808-815
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• 2017

We demonstrated the quantitative detection of a toxin-producing Microcystis aeruginosa (M. aeruginosa) strain with the laboratory protocol of the NanoGene assay. The NanoGene assay was selected because its laboratory protocol is in the process of being transplanted into a portable system. The mcyD gene of M. aeruginosa was targeted and, as expected, its corresponding fluorescence signal was linearly proportional to the mcyD gene copy number. The sensitivity of the NanoGene assay for this purpose was validated using both dsDNA mcyD gene amplicons and genomic DNAs (gDNA). The limit of detection was determined to be 38 mcyD gene copies per reaction and 9 algal cells/ml water. The specificity of the assay was also demonstrated by the addition of gDNA extracted from environmental algae into the hybridization reaction. Detection of M. aeruginosa was performed in the environmental samples with environmentally relevant sensitivity (${\sim}10^5$ algal cells/ml) and specificity. As expected, M. aeruginosa were not detected in nonspecific environmental algal gDNA over the range of $2{\times}10^0$ to $2{\times}10^7$ algal cells/ml.

• Fisheries and Aquatic Sciences
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• v.16 no.4
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• pp.273-277
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• 2013

Vibriosis caused by Vibrio anguillarum and edwardsiellosis caused by Edwardsiella tarda are septicemic diseases of many commercially important freshwater and marine fishes, and threaten the aquaculture industry in Korea. Early diagnosis and accurate identification of these two bacterial species could help to prevent these diseases and minimize the damage to cultured marine species. This study designed a duplex polymerase chain reaction (PCR) method for the simultaneous detection of two major fish pathogens: V. anguillarum and E. tarda. Each pair of oligonucleotide primers exclusively amplified the target groEL gene of the specific microorganism. Twenty-two Vibrio and ten non-Vibrio enteric species were used to check the specificity of the primers, which were found to be highly specific for the target species, even among closely related species. The detection limit was 400 pg for V. anguillarum and 4 ng for E. tarda when mixed purified DNA was used as the template. This assay showed high specificity and sensitivity in the simultaneous detection of V. anguillarum and E. tarda from artificially inoculated seawater and fish.

• Genomics & Informatics
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• v.19 no.3
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• pp.30.1-30.8
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• 2021

Salmonella species are among the major pathogens that cause foodborne illness outbreaks. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the rapid and sensitive detection of Salmonella species. We designed LAMP primers targeting the hilA gene as a universal marker of Salmonella species. A total of seven Salmonella species strains and 11 non-Salmonella pathogen strains from eight different genera were used in this study. All Salmonella strains showed positive amplification signals with the Salmonella LAMP assay; however, there was no non-specific amplification signal for the non-Salmonella strains. The detection limit was 100 femtograms (20 copies per reaction), which was ~1,000 times more sensitive than the detection limits of the conventional polymerase chain reaction (PCR) assay (100 pg). The reaction time for a positive amplification signal was less than 20 minutes, which was less than one-third the time taken while using conventional PCR. In conclusion, our Salmonella LAMP assay accurately detected Salmonella species with a higher degree of sensitivity and greater rapidity than the conventional PCR assay, and it may be suitable for point-of-care testing in the field.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.112-116
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• 2003

An immunosensor based on surface plasmon resonance (SPR) onto a protein G layer by Self-assembly technique was developed for detection of Legionella pneumophila. The protein G layer by self-assembly technique was fabricated on a gold (Au) surface by adsorbing the 11-mercaptoundecanoic acid (MUA) and an activation process for the chemical binding of the free amino (-NH$_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) in series. The formation of the protein G layer by self-assembly technique on the Au Substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The Surface topographies of the fabricated thin films on an Au substrate were also analyzed by using an atomic force microscope (AFM). Consequently, an immunosensor for the detection of L. pneumophila using SPR was developed with a detection limit of up to 10$^2$CFU per mL.

• Asian Journal of Atmospheric Environment
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• v.7 no.2
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• pp.105-113
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• 2013

We constructed and tested an airborne peroxyacetyl nitrate (PAN) monitoring system based on luminol chemiluminescence detection with fast gas chromatography. This system allowed for simultaneous measurement of PAN and nitrogen dioxide ($NO_2$) with a time resolution of <2 min. Actual sample masses within the fixed volume sample loop at various altitudes and temperatures were adjusted to standard atmosphere, using measured pressures and temperatures. The airborne PAN measurement system was evaluated during two field studies above the southern Korean Peninsula in August and October 2009. The detection limit based on the ISO approach was 0.035 ppbv PAN, well below the observed concentrations of 0.185-1.49 ppbv during these studies. Under these conditions, the PAN mixing ratios were positively correlated with $O_x$ ($O_x=O_3+NO_2$), with slopes varying between 0.014 and 0.033 and intercepts between 22.6 and 55.1 ppbv $O_x$. The intercepts corresponded roughly to background $O_x$ mixing ratios in central Europe; however, the slopes were above the range of slopes reported in other studies. We also enhanced the durability, safety, and ease of maintenance of the PAN monitoring system by redesigning the structure of the conventional luminol cell.

• YAKHAK HOEJI
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• v.40 no.1
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• pp.41-45
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• 1996

A high performance liquid chromatography using photoreduction fluorescence detection was described for the analysis of saikosaponins. Saikosaponins were separated on an $NH_2$ column using acetonitrile and aqueous 2-tert-butylanthraquinone(t-BAQ) as mobile phase. Column effluent was passed through a 40cm PTFE capillary tube coiled around a 10W UV lamp to reduce t-BAQ to a highly fluorescent dihydroxyanthracene derivative which was detected by a fluorescence detector. The optimal concentration of t-BAQ was found to be $6{\times}10^{-5}M$ and the optimal reaction time to be 2 seconds. The detection limit for saikosaponin a and d by this method was found to be about 280ng and 80ng. The dynamic linear range was over two orders and the correlation coefficient of the calibration curve of them was 0.998.

• Smart Structures and Systems
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• v.12 no.3_4
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• pp.309-325
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• 2013

Environmental and operational variables are inevitable concerns by researchers and engineers when implementing the damage detection algorithm in practical projects, because the change of structural behavior could be masked by the conditions in a large extent. Thus, reliable damage detection methods should have a virtue of immunity from environmental and operational variables. In this paper, the pair cointegration method was presented as a novel way to remove the effect of environmental variables. At the beginning, the concept and procedure of this approach were introduced, and then the theoretical formulation and numerical simulations were put forward to illustrate the feasibility. The jump exceeding the control limit in the residual indicates the occurrence of damage, while the direction and magnitude imply the most potential damage location. In addition, the simulation results show that the proposed method has strong ability to resist the noise.

• BioChip Journal
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• v.12 no.4
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• pp.326-331
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• 2018

Contamination by pesticides is an everincreasing problem associated with fields of environmental management and healthcare. Accordingly, appropriate treatments are in demand. Pesticide detection methods have been researched extensively, aimed at making the detection convenient, fast, cost-effective, and easy to use. Among the various detecting strategies, paper-based assay is potent for real-time pesticide sensing due to its unique advantages including disposability, light weight, and low cost. In this study, a paper-based sensor for chlorpyrifos, an organophosphate pesticide, has been developed by layering three sheets of patterned plates. In colorimetric quantification of pesticides, the blue color produced by the interaction between acetylcholinesterase and indoxyl acetate is inhibited by the pesticide molecules present in the sample solutions. With the optimized paper-based sensor, the pesticide is sensitively detected (limit of detection =8.60 ppm) within 5min. Furthermore, the shelf life of the device is enhanced to 14 days after from the fabrication, by treating trehalose solution onto the deposited reagents. We expect the paper-based device to be utilized as a first-screening analytic device for water quality monitoring and food analysis.