• Title/Summary/Keyword: Detection agent

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Manufacturing Protein-DNA Chip for Depigmenting Agent Screening (전사인자 저해제 통한 미백제 탐색용 단백질 칩 제작)

  • Han Jung-Sun;Kwak Eun-Young;Lee Hyang-Bok;Shin Jlung-Hyun;Baek Seung-Hak;Chung Bong-Hyun;Kim Eun-Ki
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.479-483
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    • 2004
  • An attempt was made to develop a proteinchip for screening of MITF (microphthalmia transcription factor) inhibitor. Binding of MITF to E-box causes transcription of several pigmenting genes including tyrosinase gene. We investigated binding of MITF and its DNA binding site (E-box) using a protein-DNA chip with various detection methods including flurorescence (Cyt3). SPR (surface plasmon resonance) and SPRi (surface plasmon resonance imaging). A fusion protein (MITF-Maltose Binding Protein) was attached on the glass plate by chemical modification. An inhibitory synthetic DNA oligomer, artificially designed based on the E-box sequence, inhibited the binding of MITF and E-box. These results showed the potentials of flurorescence-based MITF protein chip as a microarray for high throughput screening (HTS) system of depigmenting agents.

Molecular Identification and Real-time Quantitative PCR (qPCR) for Rapid Detection of Thelohanellus kitauei, a Myxozoan Parasite Causing Intestinal Giant Cystic Disease in the Israel Carp

  • Seo, Jung-Soo;Jeon, Eun-Ji;Kim, Moo-Sang;Woo, Sung-Ho;Kim, Jin-Do;Jung, Sung-Hee;Park, Myoung-Ae;Jee, Bo-Young;Kim, Jin-Woo;Kim, Yi-Cheong;Lee, Eun-Hye
    • Parasites, Hosts and Diseases
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    • v.50 no.2
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    • pp.103-111
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    • 2012
  • Intestinal giant-cystic disease (IGCD) of the Israel carp (Cyprinus carpio nudus) has been recognized as one of the most serious diseases afflicting inland farmed fish in the Republic of Korea, and Thelohanellus kitauei has been identified as the causative agent of the disease. Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations. However, these types of diagnostic examinations are relatively time-consuming, and the infection frequently cannot be detected in its early stages. In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans. Moreover, conventional PCR and real-time quantitative PCR (qPCR) using oligonucleotide primers for the amplification of 18S rRNA gene fragment were established for further use as methods for rapid diagnosis of IGCD. Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.

Specific and Sensitive Detection of Venturia nashicola, the Scab Fungus of Asian Pears, by Nested PCR

  • Koh, Hyun Seok;Sohn, San Ho;Lee, Young Sun;Koh, Young Jin;Song, Jang Hoon;Jung, Jae Sung
    • The Plant Pathology Journal
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    • v.29 no.4
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    • pp.357-363
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    • 2013
  • The fungus Venturia nashicola is the causal agent of scab on Asian pears. For the rapid and reliable identification as well as sensitive detection of V. nashicola, a PCR-based technique was developed. DNA fingerprints of three closely related species, V. nashicola, V. pirina, and V. inaequalis, were obtained by random amplified polymorphic DNA (RAPD) analysis. Two RAPD markers specific to V. nashicola were identified by PCR, after which two pairs of sequence characterized amplified region (SCAR) primers were designed from the nucleotide sequences of the markers. The SCAR primer pairs, designated as D12F/D12R and E11F/E11R, amplified 535-bp and 525-bp DNA fragments, respectively, only from genomic DNA of V. nashicola. The specificity of the primer sets was tested on strains representing three species of Venturia and 20 fungal plant pathogens. The nested PCR primer pair specific to V. nashicola was developed based on the sequence of the species-specific 525-bp DNA fragment amplified by primer set E11F/E11R. The internal primer pair Na11F/Na11R amplified a 235-bp fragment from V. nashicola, but not from any other fungal species tested. The nested PCR assay was sensitive enough to detect the specific fragment in 50 fg of V. nashicola DNA.

Discovery of antigens for early detection of Mycobacterium avium subsp. paratuberculosis and analysis of characteristics using bioinformatics tools (Mycobacterium avium subsp. paratuberculosis 감염 초기 개체 검출을 위한 항원 탐색 및 특성 분석)

  • Park, Hong-Tae;Park, Hyun-Eui;Shin, Min-Kyoung;Cho, Yong-Il;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.55 no.2
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    • pp.89-95
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    • 2015
  • Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is one of the most widespread and economically important diseases in cattle. Current diagnostic methods are based on the detection of anti-MAP antibodies in serum or isolation of the causative agent. However, these techniques are often not applicable for cases of subclinical infection due to relatively low sensitivity. Therefore, finding new antigen candidates that strongly react with the host immune system had been attempted. To effectively detect infection during the subclinical stage, several antigen candidates were selected based on previous researches. Characteristics of the selected antigen candidates were analyzed using bioinformatics-based prediction tools. A total of nine antigens were selected (MAP0862, MAP3817c, MAP2077c, MAP0860c, MAP3954, MAP3155c, MAP1204, MAP1087, and MAP2963c) that have MAP-specific and/or high immune responses to infected animals. Using a transmembrane prediction tool, five of the nine antigen candidates were predicted to be membrane protein (MAP3817c, MAP3954, MAP3155c, MAP1087, and MAP1204). Some of the predicted protein structures identified using the I-TASSER server shared similarities with known proteins found in the Protein Data Bank database (MAP0862, MAP1204, and MAP2077c). In future studies, the characteristics and diagnostic efficiency of the selected antigen candidates will be evaluated.

A Development of The Road Surface Decision Algorithm Using SVM(Support Vector Machine) Clustering Methods (SVM(Support Vector Machine) 기법을 활용한 노면상태 판별 알고리즘 개발)

  • Kim, Jong Hoon;Won, Jae Moo
    • The Journal of The Korea Institute of Intelligent Transport Systems
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    • v.12 no.5
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    • pp.1-12
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    • 2013
  • Road's accidents caused by Ice, snow, Wet of roads surface conditions and weather conditions situations that are constantly occurring. That is, driver's negligence and safe driving ability of individuals due to lack of awareness, and Road management main agent(the government and the public, etc.) due to road conditions, if there is insufficient information. So Related research needs is a trend that is required. In this study, gather Camera(Stereo camera)'s image data, and analysis polarization coefficients and wavelet transform. And unlike traditional single-dimensional classification algorithms as multi-dimensional analysis by using SVM classification techniques, develop an algorithm to determine road conditions. Four on the road conditions (dry, wet, snow, ice) recognition success rate for the detection and analysis of experiments.

First detection of avian polyomavirus by PCR from Alexandrine Parakeet (Psittacula eupatria) in Korea (대본청 앵무(Psittacula eupatria )로부터 PCR에 의한 avian polyomavirus 최초 검출)

  • Kim, Hee-Jung;Lee, Sun-Rock;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.37 no.3
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    • pp.213-218
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    • 2014
  • In early April 2014, a month-old Alexandrine Paraqeet (Psittacula eupatria) that was raised in a domestic aviary located in Gyungju-si, Korea was suddenly died and submitted to Animal Disease Intervention Center, Kyungpook National University in order to diagnose the causative agent. In post-mortem examination, the bird had abnormally developed feathers on the neck and abdomen region and subcutaneous hemorrhages on the neck and cheek adjacent to the beak. At necropsy, the bird had hemorrhage on the muscle of the femoral region, ascites, multi-focal hemorrhages on the epicardium, and diffuse hemorrhages on the sub-serosa of proventriculus and gizzard, suggesting typical avian polyomavirus (APV) infection. The partial large tumor (T) antigen gene of APV was detected by PCR from tissues of the heart, lung, liver, kidney, proventriculus and feathers of the APV-suspected birds. However, other pathogenic virus-specific nucleic acid common with psittacine birds such as avian bornavirus, psittacine beak and feather disease virus and psittacid herpesvirus were not detected from the mixed tissue samples of the bird, indicating this case is due to single infection of APV. Nucleotide sequence analysis of the partially amplified large T antigen DNA was confirmed to have 99~100% homology with that of the previously reported APV strains. This case report describes the first detection of APV in Alexandrine Paraqeet in Korea.

Development of competitive enzyme linked immunosorbent assay for detection of Coxiella burnetii antibody in animal (동물에서 Coxiella burnetii 항체를 진단하기 위한 경쟁효소면역법 개발)

  • Cho, Dong-hee;Kim, Yong-ju;Wee, Sung-hwan;Cho, Mi-young;Kweon, Chang-hee;Kang, Yung-bai;Park, Yong-ho;Cho, Sang-nae
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.81-85
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    • 2000
  • Coxiella burnetii (C burnetii) is the causative agent of Q fever in animal and human. The distribution of the disease has been documented around world. In this study we developed the competitive enzyme linked immunosorbent assay(cELISA) and compared it with indirect immunofluorescent assay(IFA). A monoclonal antibody(Mab) against C burnetii and a peroxidase-conjugated anti-mouse IgM were used as an indicator system competing against antibody in animal serum or as an indicater of the absence of antibody. Sera were considered antibody positive when the percentage inhibition index(PI index) is upper than 30. PI index is calculated as 100-[sample OD/Mab OD)${\times}100$]. Among 162 bovine serum samples, 23 samples were antibody positive both in cELISA and IFA. And 156 samples showed same results. From goat with experimentally induced infection with C burnetii the antibody was detected 20 days early in cELISA compared to IFA. On the basis of present findings, it was demonstrated that cELISA is a reliable diagnostic method for The detection of specific antibodies against C burnetii infection.

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Application of fluorescent particles as a tracer to detect the membrane surface damage in a pilot scale membrane bioreactor (형광입자를 이용한 분리막 표면 검측 방법의 파일럿 규모 플랜트 적용)

  • Kim, Choah;Kim, Hee Jun;Cho, Jinwoo
    • Journal of Korean Society of Water and Wastewater
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    • v.30 no.1
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    • pp.33-40
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    • 2016
  • In this study, a fluorescent silica nano particle is used as the surrogate for challenging test of membrane surface integrity. The particles are functionalized by a fluorescent dying agent so that as an ultraviolet light is imposed a bright fluorescent image from the particles can be taken. If a membrane surface is damaged and has a compromised part larger than the size of surrogate the fluorescent particles would pass through and contained in the permeate. An operator can directly notice whether the membrane surface is damaged or not by detecting a fluorescent image taken from the permeate. Additionally, the size of compromised part is estimated through analysing the fluorescent image in which we surmise the mass of particles included in the permeate by calculating an average RGB value of the image. The pilot scale experiments showed that this method could be applied successfully to determine if a membrane surface had a damaged parts regardless of the test condition. In the testing on the actual damaged area of $4.712mm^2$, the lowest error of estimating the damaged area was -1.32% with the surrogate concentration of 80 mg/L, flux of $40L/m^2/hr$ for 25 minutes of detection. A further study is still going on to increase the lowest detection limit and thus decrease the error of estimation.

Detection and Molecular Identification of Human Enteric Viruses in Urban Rivers in Korea

  • Lee, Cheong-Hoon;Kim, Sang-Jong
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.171-171
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    • 2008
  • We monitored the occurrence of human enteric viruses in urban rivers by cell culture-PCR and RT-nested PCR. Water samples were collected monthly or semimonthly between May 2002 and March 2003 in four urban tributaries. Enteric viruses were detected by RT-nested PCR and cell culture-PCR based on a combination of Buffalo Green monkey kidney (BGMK) and A549 cell lines, followed by phylogenetic analysis of amplicons. By RT-nested PCR analysis, 45 (77.6%), 32 (55.2%), 32 (55.2%), 26 (44.8%), 12 (20.7%), 2 (3.4%), 4 (6.9%), and 4 (6.9%) of 58 samples showed positive results with adenoviruses, enteroviruses, noroviruses (NV) genogroup I (GI) and II (GII), reoviruses, hepatitis A viruses, rotaviruses and sapoviruses, respectively. Adenoviruses were most often detected and only eight (13.8%) samples were negative for adenoviruses and positive for other enteric viruses in the studied sites. Thirty-one (77.5%) of the 40 samples were positive for infectious adenoviruses and/or enteroviruses based on cell culture-PCR, and the frequency of positive samples grown on A549 and BGMK (65.0%) was higher than that grown on BGMK alone (47.5%). The occurrence of each enteric virus, except reoviruses and hepatitis A viruses was not statistically correlated with the water temperature and levels of fecal coliforms according to Binary logistic regression model. By sequence analysis, most strains of adenoviruses and enteroviruses detected in this study are similar to the causative agent of viral diseases in Korea and most NV GI- and GII-grouped strains were closely related to the reference strains from China and Japan, and GII/4-related strains had similar sequences to strains recognized as a worldwide epidemic outbreak. Our results suggested that monitoring human enteric viruses is necessary to improve microbial quality and cell culture-PCR using the combination of A549 and BGMK cells and the adenovirus detection by PCR could be useful for monitoring viral contamination in the aquatic environment.

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The comparison of pathogenicity in the SPF chickens challenged with avian reovirus and/or fowl adenovirus (SPF 닭에서 레오바이러스와 아데노바이러스의 단일 혹은 혼합감염에 따른 병원성 비교)

  • Min, Kyeong-Cheol;Choi, Dong-Myong;Kim, Ji-Yea;Jeon, Eun-Ok;Byun, Sung-Hwan;Mo, In-Pil
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.139-148
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    • 2011
  • Avian reovirus (ARV) and fowl adenovirus (FAdV) were evaluated for pathogenicity in specific pathogen free (SPF) chickens. ARV was isolated from the broilers with history of malabsorption syndrome (MAS). FAdV was isolated from the layer breeders with inclusion body hepatitis and hydropericardium syndrome. Total 6 inoculated groups including 1 un-inoculated group were organized and inoculated with the ARV and/or FAdV by oral route. The minimal pathological lesions and lower viral gene detection rates were present in the ARV inoculated groups compared to those of FAdV or ARV/FAdV inoculated groups. Common gross lesions in the ARV inoculated group were distended intestine with foamy contents and in the FAdV group there were foamy cecal contents and hydropericardium among the evaluation methods such as gross and histological lesion, viral gene detection, body weight and serum chemistry, histopathological lesion score was reliable especially in the liver lesions such as hepatic necrosis and lymphocytic infiltration. However, we did not success to evaluate the synergetic effect of mixed infection of ARV and FAdV in this study. Therefore, we need further study to reproduce malabsorption syndrome of ARV infection using different viral agent such as rotavirus and using different dose of virus.