• 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.

• Journal of the Korean Society for Heat Treatment
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• v.34 no.6
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• pp.287-293
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• 2021

This study was intended to investigate the influence of Al content on hardness and microstructural characteristics of discontinuous precipitates (DPs) formed by isothermal aging in Mg-8.7%Al and Mg-10%Al alloys. In order to obtain large amount of DPs in the microstructure, the alloy specimens were solution-treated at 688K for 24 h followed by water quenching, and then aged at 418K for 48h. The Mg-Al alloy with higher Al content was characterized by higher volume fraction of DPs at the same aging condition, lower interlamellar spacing of the DPs, thinner β phase layer and higher β phase content in the DPs. This is closely related to the higher velocity of discontinuous precipitation process resulting from the higher Al supersaturation in the α-(Mg) matrix. The Mg-10%Al alloy showed higher hardness of the DPs and greater difference in hardness between as-cast state and DPs than the Mg-8.7%Al alloy.

• Research in Plant Disease
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• v.28 no.2
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• pp.92-97
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• 2022

Apple stem grooving virus (ASGV) is a high-risk viral pathogen that infects many types of fruit trees, especially pear and apple, and causes serious economic losses across the globe. Thus, rapid and reliable detection assay is needed to identify ASGV infection and prevent its spread. A reliable reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed, optimize, and evaluated for the coding region of coat protein of ASGV in pear leaf. The developed RT-LAMP facilitated the simple screening of ASGV using visible fluorescence and electrophoresis. The optimized reaction conditions for the RT-LAMP were 63℃ for 50 min, and the results showed high specificity and 100-fold greater sensitivity than the reverse transcription polymerase chain reaction. In addition, the reliability of the RT-LAMP was validated using field-collected pear leaves. Furthermore, the potential application of paper-based RNA isolation, combined with RT-LAMP, was also evaluated for detecting ASGV from field-collected samples. These assays could be widely applied to ASGV detection in field conditions and to virus-free certification programs.

• Journal of the Korean Society for Heat Treatment
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• v.36 no.5
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• pp.277-284
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• 2023

Hardness and damping characteristics of fine discontinuous precipitates (DPs) microstructure generated by low temperature long term isothermal aging were investigated in comparison with those of T6 heat-treated microstructure composed of DPs and continuous precipitates (CPs) in Mg-9%Al alloy. In this study, T6 and fine DPs microstructures were obtained by isothermal aging at 453 K for 24 h and at 413 K for 336 h, respectively, after solution treatment at 693 K for 24 h. The DPs microstructure exhibited higher hardness than the T6 microstructure, which is related to the lower (α + β) interlamellar spacing of the DPs. The DPs microstructure possessed better damping capacity than the T6 microstructure in the strain-amplitude independent region, whereas in the strain-amplitude dependent region, the reverse behavior was observed. The damping tendencies depending on strain-amplitude were discussed based on the microstructural features of the T6 and DPs microstructures.

• Genomics & Informatics
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• v.20 no.4
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• pp.46.1-46.7
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• 2022

Influenza A virus (IAV) is the most widespread pathogen causing human respiratory infections. Although polymerase chain reaction (PCR)-based methods are currently the most commonly used tools for IAV detection, PCR is not ideal for point-of-care testing. In this study, we aimed to develop a more rapid and sensitive method than PCR-based tools to detect IAV using loop-mediated isothermal amplification (LAMP) technology. We designed reverse-transcriptional (RT)-LAMP primers targeting the hemagglutinin gene. RNAs from reference H1N1 and H3N2 showed specific RT-LAMP signals with the designed primers. We optimized the reaction conditions and developed universal reaction conditions for both LAMP assays. Under these conditions, the detection limit was 50 copies for both RT-LAMP assays. There was no non-specific signal to 19 non-IAV respiratory viruses, such as influenza B virus, coronaviruses, and respiratory syncytial viruses. Regarding the reaction time, a positive signal was detected within 25 min after starting the reaction. In conclusion, our RT-LAMP assay has high sensitivity and specificity for the detection of the H1 and H3 subtypes, making it suitable for point-of-care IAV testing.

• Journal of Life Science
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• v.26 no.5
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• pp.620-631
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• 2016

Viroids are about 250-400 base pair of short single strand RNA fragments have been associated with economically important plant diseases. Due to the lack of protein expression capacity associated with replication, it is very difficult to diagnosis viroid diseases in serological methods. For detecting viroid at plants, molecular-based techniques such as agarose gel electrophoresis, polyacrylamide gel electrophoresis (PAGE), DNA-hybridization, blotting analysis and conventional RT-PCR are reliable. Real-time RT-PCR methods that grafted on RT-PCR methods with improved confirmation methods have been also utilized. However, they are still labor-intensive, time-consuming, and require personnel with expertise. Loop-mediated Isothermal Amplification (LAMP) method is a nucleic acid amplification method under the isothermal condition. The LAMP methodology has been reported to be simple, rapid, sensitive and field applicable in detecting a variety of pathogens. The results of LAMP method can be colorized by adding a visible material such as SYBR green I, Evagreen, Calcein, Berberine and Hydroxy naphthol blue (HNB) with simple equipment or naked eyes. The combination of LAMP method and nucleic pathogens, viroids, can be used to realize simple diagnosis platform for the genetic point-of care testing system. The aim at this review is to summary viroid-caused diseases and the simple visible approach for diagnosing viroids using Loop-mediated Isothermal Amplification (LAMP) method.

• Research in Plant Disease
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• v.21 no.4
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• pp.315-320
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• 2015

Soybean mosaic virus (SMV) is a prevalent pathogen that causes significant yield reduction in soybean production worldwide. SMV belongs to potyvirus and causes typical symptoms such as mild mosaic, mosaic and necrosis. SMV is seed-borne and also transmitted by aphid. Eleven SMV strains, G1 to G7, G5H, G6H, G7H, and G7a were reported in soybean varieties in Korea. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method allowed one-step detection of gene amplification by simple procedure and needed only a simple incubator for isothermal template. This RT-LAMP method allowed direct detection of RNA from virus-infected plants without thermal cycling and gel electrophoresis. In this study, we designed RT-LAMP primers named SML-F3/B3/FIP/BIP from coat protein gene sequence of SMV. After the reaction of RT-LAMP, products were identified by electrophoresis and with the detective fluorescent dye, SYBR Green I under daylight and UV light. Optimal reaction condition was at $58^{\circ}C$ for 60 min and the primers of RT-LAMP showed the specificity for nine SMV strains tested in this study.

• Polymer(Korea)
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• v.25 no.3
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• pp.359-366
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• 2001

Isothermal physical aging of a glass fiber/epoxy composite was examined at different aging temperatures ($T_a$) and degrees of conversion (monitored by the glass transition temperature, $T_g$) by means of the TBA torsion pendulum technique. The range of aging temperature was from 10 to $130^{\circ}C$ : the conversion was systematically changed from $T_g$=$76^{\circ}C$ to $T_g$=$177^{\circ}C$ (fully crosslinked). The effect of isothermal physical aging was manifested as perturbations of the modulus and mechanical loss vs. temperature in the vicinity of $T_a$ for all conversions. The rate of isothermal physical aging determined from the change of modulus with aging time at fixed aging temperature decreased and then increased with increasing conversion below T$_{a}$=9$0^{\circ}C$. There exists a superposition in aging rate vs. ($T_g$ -$T_a$) by shifting horizontally and vertically. This implies that the physical aging process is independent of the change of chemical structure as conversion proceeds. It has been found that water absorbed at the aging temperature below $70^{\circ}C$ during isothermal physical aging lowers the apparent aging rate. It is due to the absorbed water molecules forming strong polar interactions with hydroxyl group on network chain and reducing the segmental mobility during the physical aging.g.

• Proceedings of the Materials Research Society of Korea Conference
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• 1994.11a
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• pp.167-167
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• 1994

• Proceedings of the Korean Fiber Society Conference
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• 1999.10a
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• pp.9-12
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• 1999