• Journal of Plant Biotechnology
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• 제47권4호
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• pp.309-315
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• 2020

Advances in biotechnology have led to progress in crop genetic engineering to improve agricultural productivity. The use of genetically modified (GM) crops has increased, as have consumers' and regulators' concerns about the safety of GM crops to human health, and ecological biodiversity. As such, the identification of GM crops is a critical issue for developers and distributors, and their labeling is mandatory. Multiplex polymerase chain reaction (PCR) has been developed and its use validated for the detection and identification of GM crops in quarantine. Herein, we established a simultaneous detection method to identify four GM maize events. Event-specific primers were designed between the junction region of transgene and genome of four GM maize lines, namely 5307, DAS-40278-9, MON87460, and MON87427. To verify the efficiency and accuracy of the multiplex PCR we used specificity analysis, limit of detection evaluation, and mixed certified reference materials identification. The multiplex PCR method was applied to analyze 29 living, modified maize volunteers collected in South Korea in 2018 and 2019. We performed multiplex PCR analysis to identify events and confirmed the result by simplex PCR using each event-specific primer. As a result, rather than detecting each event individually, the simultaneous detection PCR method enabled the rapid analysis of 29 GM maize volunteers. Thus, the novel multiplex PCR method is applicable for living modified organism volunteer identification.

• Journal of Veterinary Science
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• 제23권5호
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• pp.32.1-32.11
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• 2022

Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases. The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions: The spbELISA has practical applications in assessing the vaccination status of large pig herds.

• 대한수의학회지
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• 제44권4호
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• pp.655-662
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• 2004

The aim of this work was the validation of a rapid real-time PCR assay based on TaqMan technology for the unequivocal identification of infectious canine hepatitis (ICH) virus, to be used directly on DNA purified from blood specimens. A real-time PCR system targeting at the E3 ORFA gene sequence of canine adenovirus type 1 was optimized and validated through comparative analysis of samples using conventional PCR system. The real-time PCR assay based on TaqMan technology could disclose 23 (37.7%) out of 61 samples as PCR positive. In contrast, 18 (29.5%) samples were found PCR positive when conventional PCR was applied on these samples. The use of the ABI Prism 7700 sequence detection system allowed the efficient determination of the amplified product accumulation through a fluorogenic probe. The entire real-time TaqMan PCR assay, including DNA extraction, amplification, and detection could be completed within 3 hours. The detection method of real-time TaqMan PCR assay was 1,000 times more sensitive than conventional PCR. Real-time TaqMan probe and primer set developed and optimized in this study is a sensitive, rapid and accurate method for detection of ICH virus and can be effective screening tool for the detection of ICH in a diagnostic laboratory routines.

• Bulletin of the Korean Chemical Society
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• 제35권4호
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• pp.1082-1086
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• 2014

A multi-channel microchip electrophoresis (MCME) method with parallel laser-induced fluorescence (LIF) detection was developed for rapid screening of H1N1 virus. The hemagglutinin (HA) and nucleocapsid protein (NP) gene of H1N1 virus were amplified using polymerase chain reaction (PCR). The amplified PCR products of the H1N1 virus DNA (HA, 116 bp and NP, 195 bp) were simultaneously detected within 25 s in three parallel channels using an expanded laser beam and a charge-coupled device camera. The parallel separations were demonstrated using a sieving gel matrix of 0.3% poly(ethylene oxide) ($M_r$ = 8,000,000) in $1{\times}$ TBE buffer (pH 8.4) with a programmed step electric field strength (PSEFS). The method was ~20 times faster than conventional slab gel electrophoresis, without any loss of resolving power or reproducibility. The proposed MCME/PSEFS assay technique provides a simple and accurate method for fast high-throughput screening of infectious virus DNA molecules under 400 bp.

• 정보보호학회논문지
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• 제21권1호
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• pp.213-219
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• 2011

스마트폰 시장이 급속하게 성장함에 따라 보안위협도 동시에 증가하고 있다. 가장 큰 스마트폰 보안위협 중 하나는 스마트폰 마켓에 안전성이 검증되지 않은 어플리케이션이 유통되고 있다는 점이다. 안드로이드 마켓의 경우 어플리케이션 검증을 수행하지 않아 악성 어플리케이션이 유통되고 있는 상황이다. 이와 같이 마켓을 통해 유포되는 악성 어플리케이션에 대응하기 위해서는 안드로이드 어플리케이선의 악성 행위 여부를 탐지할수 있는 기술이 필요하다. 본 논문에서는 안드로이드 어플리케이션의 악성행위를 탐지할 수 있는 분석 방법을 제안하고 구현내용을 소개하고자 한다.

• 한국식품위생안전성학회지
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• 제36권1호
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• pp.1-8
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• 2021

본 연구에서는 농약의 음성시료에서는 acetylcholinesterase와 acetylthiocholine을 반응시켜 +전하와 -전하를 가지는 thiocholine으로 분해되어 금 나노입자를 응집시켜 역 Y자 스트립상에서 청자색의 반응선(띠)을 형성하고 양성 시료에서는 생성시키지 않는 원리를 이용한 신속 농약 검출법을 개발하였다. 개발한 분석법은 유기인계 농약 말라옥손과 카바메이트계 농약 카보퓨란을 각각 10 ng/mL 수준까지 검출이 가능한 것으로 확인되었으며, 2종의 유기인계와 카바메이트계 농약(EPN, dichlorvos)에 대해 추가적으로 검출 한계를 확인한 결과에서도 10 ng/mL 수준까지 모두 검출 가능함을 확인할 수 있었다. 그러나 3종의 트리아진 계열의 농약과 각 1종의 피레스로이드, 카복사마이드, 페닐아마이드 및 유기염소계열의 농약에 대해서는 반응성이 없는 것으로 확인되어 유기인계와 카바메이트계 농약 분석에 적용이 가능한 것으로 확인되었다. 마지막으로, 임의로 오염시킨 농산물 시료를 대상으로 분석법의 회수율을 확인한 결과, 말라옥손에 대해서 96.4에서100.7%, 카보퓨란은 81에서112.7%의 회수율이 확인되어 본 연구에서 개발한 역 Y자 스트립을 농약 검출법으로 이용한다면 농산물과 농업환경 중 존재하는 유기인계 및 카바메이트계 잔류농약을 신속하게 검출할 수 있을 것으로 판단된다.

• 식물병연구
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• 제21권4호
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• pp.321-325
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• 2015

다공성 세라믹 큐브를 이용한 RT-PCR 진단법은 별도의 핵산 추출 과정이나 용액 처리 없이, 식물체에 접촉시켜 큐브의 공극에 바이러스 입자나 핵산 등의 분자가 신속하게 흡수되면 이를 바로 RT-PCR 반응에 넣어 유전자를 증폭시키는 방법으로, 식물체로부터 빠르고 정확하게 바이러스를 진단하는 방법이다. 본 연구에서는 다공성 세라믹 큐브를 이용하여 벼에 발생하는 주요 바이러스인 벼줄무늬잎마름바이러스(RSV)를 진단하는 RT-PCR 진단법을 확립하였다. 벼의 잎, 잎집, 또는 줄기를 대상으로 큐브 1개 또는 3개를 사용하여 즙액을 흡수시킨 후, 이를 RT-PCR 주형으로 사용하였고, 그 결과 변성처리에 큰 차이 없이 증폭 효율이 나타났다. 또한 즙액을 흡수한 큐브는 9주차까지 상온에서 보관한 후 RT-PCR을 실시하여도 안정적으로 증폭 효율을 나타내었다.

• 식물병연구
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• 제7권2호
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• pp.80-85
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• 2001

이 연구는 고랭지 나리에서 발생하는 바이러스의 병징, 종류 및 계통별 방병률을 조사 분석하고 효과적인 검정방법을 개발하고자 수행하였다. 고랭지 나리에서 발생하는 바이러스의 병징은 모자이크, 축엽, 퇴록반점, 줄무의, 라인패턴을 나타내었으며, 증상별 분포는 모자이크가 43.8%, 축엽이 29.2%, 퇴록반점이 10.9%였다. 바이러스 종류별로는 Lily symptomless virus(LSV), Cucumber mosaic virus(CMV), Lily mottle virus(LMoV) 등 6가지 바이러스가 전자현미경으로 검정되었다. 지역별로는 강릉(왕산)이 대관령보다 바이러스 이병률이 높았으며, 계통별 바이러스 이병률은 오리엔탈 계통(카사블랑카, 마르코폴로)이 아시아틱 계통(솔레미오, 플라토)보다 2~4배 높았다. 바이러스 진단방법으로는 기존의 PT-PCR보다 개선된 one-step RT-PCR 검정이 시간을 줄이면서 민감도가 뛰어나 가장 효과적이었다.

• Parasites, Hosts and Diseases
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• 제59권1호
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• pp.15-22
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• 2021

Artemisinin resistance (ART) has been confirmed in Greater Mekong Sub-region countries. Currently, C580Y mutation on Pfkelch13 gene is known as the molecular marker for the detection of ART. Rapid and accurate detection of ART in field study is essential to guide malaria containment and elimination interventions. A simple method for collection of malaria-infected blood is to spot the blood on filter paper and is fast and easy for transportation and storage in the field study. This study aims to evaluate LAMP-SNP assay for C580Y mutation detection by introducing an extra mismatched nucleotide at the 3' end of the FIP primer. The LAMP-SNP assay was performed in a water bath held at a temperature of 56℃ for 45 min. LAMP-SNP products were interpreted by both gel-electrophoresis and HNB-visualized changes in color. The method was then tested with 120 P. falciparum DNA from dried blood spot samples. In comparing the LAMP-SNP assay results with those from DNA sequencing of the clinical samples, the 2 results fully agreed to detect C580Y. The sensitivity and specificity of the LAMP-SNP assay showed 100%. There were no cross-reactions with other Plasmodium species and other Pfkelch13 mutations. The LAMP-SNP assay performed in this study was rapid, reliable, and useful in detecting artemisinin resistance in the field study.

• 대한의용생체공학회:학술대회논문집
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• 대한의용생체공학회 1998년도 추계학술대회
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• pp.201-202
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• 1998

In this paper we suggested an automated method for detecting and counting rapid eye movement(REM) using EOG during sleep. This method is formulated by two step fuzzy logic. At first step, the velocity and the distance of single channel eye movement are used for the fuzzy input to get the possibility of being REM at each EOG. At second step, the two possibility values of both EOG from the first step and the correlation coefficient of both eye movements are used for the fuzzy logic input, and the output is the final possibility of being Rapid Eye Movement. We applied this algorithm to the normal and narcoleptic sleep data and compared the difference. We found the possibility that the count of REM can be a parameter that has significant physiological meanings.