• 제목/요약/키워드: Detection agent

검색결과 461건 처리시간 0.039초

A Field Deployable Real-Time Loop-Mediated Isothermal Amplification Targeting Five Copy nrdB Gene for the Detection of 'Candidatus Liberibacter asiaticus' in Citrus

  • Tirumalareddy Danda;Jong-Won Park;Kimberly L. Timmons;Mamoudou Setamou;Eliezer S. Louzada;Madhurababu Kunta
    • The Plant Pathology Journal
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    • 제39권4호
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    • pp.309-318
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    • 2023
  • Huanglongbing (HLB) is one of the most destructive diseases in citrus, which imperils the sustainability of citriculture worldwide. The presumed causal agent of HLB, 'Candidatus Liberibacter asiaticus' (CLas) is a non-culturable phloem-limited α-proteobacterium transmitted by Asian citrus psyllids (ACP, Diaphorina citri Kuwayama). A widely adopted method for HLB diagnosis is based on quantitative real-time polymerase chain reaction (qPCR). Although HLB diagnostic qPCR provides high sensitivity and good reproducibility, it is limited by time-consuming DNA preparation from plant tissue or ACP and the requirement of proper lab instruments including a thermal cycler to conduct qPCR. In an attempt to develop a quick assay that can be deployed in the field for CLas detection, we developed a real-time loop-mediated isothermal amplification (rt-LAMP) assay by targeting the CLas five copy nrdB gene. The rt-LAMP assay using various plant sample types and psyllids successfully detected the nrdB target as low as ~2.6 Log10 copies. Although the rt-LAMP assay was less sensitive than laboratory-based qPCR (detection limit ~10 copies), the data obtained with citrus leaf and bark and ACP showed that the rt-LAMP assay has >96% CLas detection rate, compared to that of laboratory-based qPCR. However, the CLas detection rate in fibrous roots was significantly decreased compared to qPCR due to low CLas titer in some root DNA sample. We also demonstrated that the rt-LAMP assay can be used with a crude leaf DNA extract which is fully deployable in the field for quick and reliable HLB screening.

박과 작물에 과일썩음병을 일으키는 Acidovorax citrulli 검출을 위한 nested-PCR 검사법 개발 (Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae)

  • 김영탁;박경수;김혜성;이혁인;차재순
    • 식물병연구
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    • 제21권2호
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    • pp.74-81
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    • 2015
  • 박과 작물에서 과일썩음병(bacterial fruit blotch)을 일으키는 Acidovorax citrulli를 종자로부터 검출하기 위한 특이적이고 민감한 nested-PCR 방법을 개발하였다. 본 연구에서는 Next Generation Sequencing을 이용하여 draft genome sequencing을 얻은 후 이를 분석하여 PCR 프라이머를 디자인하였고, 이들 프라이머의 A. citrulli에 대한 특이성을 확인하여 Ac-ORF 21F/Ac-ORF 21R의 nested PCR 프라이머를 최종 선발하였다. Ac-ORF 21F/Ac-ORF 21R는 오직 A. citrulli에서만 특이적으로 140bp 크기의 DNA를 증폭하였으며, 그 검출민감도는 1차 PCR 검출한계(10 ng genomic DNA/PCR)보다 검출한계를 10,000배 증가시켰다. 개발된 nested-PCR 방법을 통해 병원균을 인공접종한 수박 종자의 외부검사에서 $10^1cfu/ml$까지 인공 접종 한 모든 종자 시료에서 병원균을 검출하였고, 병원균을 인공접종 한 수박 종자의 내부검사에서는 병원균이 검출되지 않았다. 자연 감염 수박 종자의 외부검사에서는 10개의 반복 시료 중 2개에서, 그리고 종자 내부검사에서는 10개의 반복 시료 중 5개에서 A. citrulli를 검출하였다. 본 연구에서 개발한 nested-PCR은 특이성과 민감도가 높고 인공접종과 자연감염 수박 종자에서도 병원균의 검출이 가능하여 박과 작물의 종자로부터 A. citrulli를 검출하는데 효과적으로 사용될 수 있을 것으로 생각된다.

친환경성 석회 도료의 개발 및 특성 연구 (Development and Characterizations of Environment-friendly Lime Paint)

  • 황대주;김호성;이승관;최문관;김환;이종대
    • 한국세라믹학회지
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    • 제46권1호
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    • pp.47-52
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    • 2009
  • Lime paint surpassing others in execution efficiency, anti-bacterial, anti-mold and small quantity emission of VOCs(Volatile Organic Compounds) characteristics was developed using a limestone as raw materials. The lime paint prepared by mixing slaked lime($37{\sim}40\;wt%$), PVA:EVA(9 wt%:1 wt%), talc(23 wt%), $TiO_2$(14 wt%), zeolite (3 wt%), antifoaming agent(5 wt%), wetting agent (5 wt%) was indicated over 99.8% of anti-bacterial and anti-mold characteristics. Also, the environment-friendly function of the lime paint was confirmed by detection of small amount of TVOCs($0.01\;mg/m^2h$) and formaldehyde($0.008\;mg/m^2h$). Execution efficiency, economy-and environment-friendly characteristics of this lime paint can make up for defects of established paints. And, it also presents the advantage of a limestone as high value added materials.

Simultaneous Determination of the Novel Neuroprotective Agent KR-31378 and its Metabolite KR-31612 Using High Performance Liquid Chromatography with Tandem Mass Spectrometry in Human Plasma

  • Kim, John;Ji, Hye-Young;Yoo, Sung-Eun;Kim, Sun-Ok;Lee, Dong-Ha;Lim, Hong;Lee, Hye-Suk
    • Archives of Pharmacal Research
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    • 제25권5호
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    • pp.647-651
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    • 2002
  • An LC/MS/MS method for the simultaneous determination of a neuroprotective agent for ischemia-reperfusion damage, KR-31378 and its N-acetyl metabolite KR-31612 in human plasma was developed. KR-31378, KR-31612 and the internal standard. KR-31543 were extracted from human plasma by liquid-liquid extraction. A reverse-phase HPLC separation was performed on Luna phenylhexyl column with the mixture of acetonitrile-5 mM ammonium formate (55:45, v/v) as mobile phase. The detection of analytes was performed using an electrospray ionization tandem mass spectrometry in the multiple reaction monitoring mode. The lower limits of quantification for KR-31378 and KR-31612 were 2.0 ng/ml. The method showed a satisfactory sensitivity, precision, accuracy, recovery and selectivity.

IP Agent 기반 IPv6 주소 관리 시스템 (IPv6 Address Management System Based on IP Agents)

  • 이동일;홍윤환;김명균
    • 한국정보통신학회:학술대회논문집
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    • 한국정보통신학회 2013년도 추계학술대회
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    • pp.1051-1054
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    • 2013
  • 기존의 IPv4주소의 한계성 때문에 전세계적으로 IPv6의 도입이 점점 가속화되고 있다. IPv6 시스템에서는 자동주소설정 기능으로 인해 각 호스트가 주소를 자동적으로 얻을 수 있다. 하지만 어떤 허가되지 않은 사용자가 악의적인 목적으로 주소를 얻고 접근할 수 있는 가능성도 존재하기 때문에 IP 주소에 대한 관리가 더욱더 중요시된다. 이를 위해서 IPv6 환경에서 인가된 호스트를 관리, 인가되지 않은 호스트의 접근을 차단하고 특정한 웹페이지로 유도하여 시스템의 보안을 유지하는 IP Agent 기반 IPv6 주소 관리 시스템을 제안한다.

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State of the Art of Anti-Screen Capture Protection Techniques

  • Lee, Young;Hahn, SangGeun
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • 제15권5호
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    • pp.1871-1890
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    • 2021
  • The transition toward a contactless society has been rapidly progressing owing to the recent COVID-19 pandemic. As a result, the IT environment of organizations and enterprises is changing rapidly; in particular, data security is expanding to the private sector. To adapt to these changes, organizations and companies have started to securely transfer confidential data to residential PCs and personally owned devices of employees working from home or from other locations. Therefore, organizations and companies are introducing streaming data services, such as the virtual desktop infrastructure (VDI) or cloud services, to securely connect internal and external networks. These methods have the advantage of providing data without the need to download to a third terminal; however, while the data are being streamed, attacks such as screen shooting or capturing are performed. Therefore, there is an increasing interest in prevention techniques against screen capture threats that may occur in a contactless environment. In this study, we analyze possible screen capture methods in a PC and a mobile phone environment and present techniques that can protect the screens against specific attack methods. The detection and defense for screen capture of PC applications on Windows OS and Mac OS could be solved with a single agent using our proposed techniques. Screen capture of mobile devices can be prevented by applying our proposed techniques on Android and iOS.

Comparative Analysis of IoT Enabled Multi Scanning Parking Model for Prediction of Available Parking Space with Existing Models

  • Anchal, Anchal;Mittal, Pooja
    • International Journal of Computer Science & Network Security
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    • 제22권8호
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    • pp.404-412
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    • 2022
  • The development in the field of the internet of things (IoT) have improved the quality of the life and also strengthened different areas in the society. All cities across the world are seeking to become smarter. The creation of a smart parking system is the essential use case in smart cities. In recent couple of years, the number of vehicles has increased significantly. As a result, it is critical to make the use of technology that enables hassle-free parking in both public and private spaces. In conventional parking systems, drivers are not able to find free parking space. Conventional systems requires more human interference in a parking lots. To manage these circumstances there is an intense need of IoT enabled parking solution that includes the well defined architecture that will contain the following components such as smart sensors, communication agreement and software solution. For implementing such a smart parking system in this paper we proposed a design of smart parking system and also compare it with convetional system. The proposed design utilizes sensors based on IoT and Data Mining techniques to handle real time management of the parking system. IoT enabled smart parking solution minimizes the human interference and also saves energy, money and time.

경남지역에서 발생한 가금티푸스의 역학적 특성 및 진단방법에 대한 비교 시험 (Epidemiological characteristics on fowl typhoid outbreak in Kyongnam province and comparison of diagnostic methods for identification of salmonella gallinarum)

  • 최유정;김도경;김용환
    • 한국동물위생학회지
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    • 제23권4호
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    • pp.349-360
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    • 2000
  • An epidemiological survey was conducted to investigate fowl typhoid outbreaks in Kyungnam province of Korea. The causative agent, salmonella gallinarum was isolated from 68 chicken samples of tentatively diagnosed fowl typhoid cases occurred during the period from January 1996 to September 1999. Comparative studies were also carried out to evaluate the diagnostic methods for detection of S gallinam The results obtained were as follows; 1. Of the 68 cases of tentatively diagnosed fowl typhoid, 56 (82%) cases were determined as fowl typhoid by biochemical test and pathological findings. The other 12 (18%) cases were determined as paratyphoid. 2. Fowl typhoid outbreaks occur continuously all seasons in the year, however the incidence was remarkably increased from May to September. 3. The frequency of incidence of fowl typhoid in terms of regional distribution was relatively high in egg-laying hens facilities, and the mode of transmission is likely to be either egg-to-egg or lateral transfer by wild birds or rats. 4. All of 18 isolates from 56 cases were identified as S gallinarum by biochemical and serological test. 5. Antimicrobial drug susceptibility test against 18 isolates showed that the isolates were highly susceptible to ASH, CZ, CF and GM (above 90%), whereas those strains were 100% resistant to EM, NA and PC. 6. S gallinarum rfbS gene was targeted to be amplified by PCR for comparative detection of S gallinarum in the experimentally infected chickens. The amplified 720bp DNA fragment, which is specific in D serogroup strains of S enterica subspecies was confirmed by agarose gel electrophoresis. 7. A comparison made between fecal culture and PCR-method revealed that later-method was relatively higher in detection rate than that of former method for S gallinarum. 8. Comparison of currently applied methods, rapid serum agglutination test (RST) and microplate agglutination test (MAT), with experimentally infected chickens were made to evaluate sensitivity of detection by neutralizing antibody titration. Both methods detected neutralizing antibodies from the challenged chickens of 5 day post infection. However, positive reactions were determined after 7 and 9 days post infection by MAT and RST, respectively.

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Simultaneous detection of fungal, bacterial, and viral pathogens in insects by multiplex PCR and capillary electrophoresis

  • Kwak, Kyu-Won;Nam, Sung-Hee;Choi, Ji-Young;Lee, Seokhyun;Kim, Hong Geun;Kim, Sung-Hyun;Park, Kwan-Ho;Han, Myung-Sae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제30권2호
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    • pp.64-74
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    • 2015
  • Beetles Protaetia brevitarsis seulensis Kolbe (Coleoptera: Cetoniidae) and Allomyrina dichotoma Linn. (Coleoptera: Scarabaeidae) are widely used in traditional medicine, and the number of insect-rearing farms is increasing in South Korea. The purpose of this study was to establish a multiplex PCR-based assay for rapid simultaneous detection of multiple pathogens causing insect diseases. Six insect parasites such as fungi Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Cordycipitaceae) and Metarhizium anisopliae (Metschn.) Sorokin (Hypocreales: Clavicipitaceae), bacteria Bacillus thuringiensis Berliner (Bacillales: Bacillaceae), Pseudomonas aeruginosa Migula (Pseudomonadales: Pseudomonadaceae), and Serratia marcescens Bizio (Enterobacteriales: Enterobacteriaceae), and Oryctes rhinoceros nudivirus were chosen based on the severity and incidence rate of insect diseases in South Korea. Pathogen-specific primers were designed and successfully applied for simultaneous detection of multiple infectious agents in farm-bred insects P. b. seulensis and A. dichotoma using multiplex PCR and high resolution capillary electrophoresis. Our results indicate that multiplex PCR is an effective and time-saving method for simultaneous detection of multiple infections in insects, and the QIAxcel capillary electrophoresis system is useful for quantitative evaluation of the individual impact of each infectious agent on the severity of insect disease. The approach designed in this study can be utilized for rapid and accurate diagnostics of infection in insect farms.

Loop-Mediated Isothermal Amplification (LAMP)법을 이용한 Pectobacterium carotovorum subsp. carotovorum의 신속 진단법 개발 (Development of a Rapid Detection Method for Pectobacterium carotovorum subsp. carotovorum Using the Loop-Mediated Isothermal Amplification (LAMP))

  • 김정구;노지나;박동석;윤병수
    • 미생물학회지
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    • 제47권2호
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    • pp.103-109
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    • 2011
  • Pectobacterium carotovorum subsp. carotovorum (PCC)는 세균성 무름병균으로 주로 감자, 양배추 등의 식물에서 질병을 일으킨다. 본 연구에서는 현장에서 신속하게 진단하기 위해 loop-mediated isothermal amplification법을 이용하여 1시간 내에 등온에서 검출 가능한 진단법을 개발하였으며, 이를 PCC-LAMP법이라 명명하였다. PCC의 lytic murein transglycolase 유전자를 특이적으로 증폭시키는 4개의 프라이머를 제작하였으며 최적 온도가 $61^{\circ}C$임을 확인하고 최적 조건을 확립하였다. 최적 조건을 바탕으로 4개의 프라이머가 $1{\times}10^3$ copies까지 검출하는 민감성을 확인할 수 있었다. 본 연구에서 개발된 PCC-LAMP법은 특이성 검사를 통해 PCC만이 특이적으로 검출됨을 확인하였으며, 이는 실제 시료에서도 적용 가능함을 확인하였다. PCC-LAMP법을 통하여 PCC를 신속하고 정확하게 검출함으로써 현장에서 유용하게 적용될 수 있을 것으로 사료된다.