• Title/Summary/Keyword: Detection agent

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Development of PCR and TaqMan PCR Assays to Detect Pseudomonas coronafaciens, a Causal Agent of Halo Blight of Oats

  • An, Ji-Hye;Noh, Young-Hee;Kim, Yong-Eon;Lee, Hyok-In;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.25-32
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    • 2015
  • Pseudomonas coronafaciens causes halo blight on oats and is a plant quarantine bacterium in many countries, including the Republic of Korea. Using of the certificated seed is important for control of the disease. Since effective detection method of P. coronafaciens is not available yet, PCR and TaqMan PCR assays for specific detection of P. coronafaciens were developed in this study. PCR primers were designed from the draft genome sequence of P. coronafaciens LMG 5060 which was obtained by the next-generation sequencing in this study. The PCR primer set Pc-12-F/Pc-12-R specifically amplified 498 bp from the 13 strains of P. coronafaciens isolated in the seven different countries (Canada, Japan, United Kingdom, Zimbabwe, Kenya, Germany, and New Zealand) and the nested primer set Pc-12-ne-F/Pc-12-ne-R specifically amplified 298 bp from those strains. The target-size PCR product was not amplified from the non-target bacteria with the PCR and nested primer sets. TaqMan PCR with Pc-12-ne-F/Pc-12-ne-R and a TaqMan probe, Pc-taqman, which were designed inside of the nested PCR amplicon, generated Ct values which in a dose-dependent manner to the amount of the target DNA and the Ct values of all the P. coronafaciens strains were above the threshold Ct value for positive detection. The TaqMan PCR generated positive Ct values from the seed extracts of the artificially inoculated oat seeds above 10 cfu/ml inoculation level. PCR and TaqMan PCR assays developed in this study will be useful tools to detect and identify the plant quarantine pathogen, P. coronafaciens.

Effect of Adjuvants on Antibody Titer of Synthetic Recombinant Light Chain of Botulinum Neurotoxin Type B and its Diagnostic Potential for Botulism

  • Jain, Swati;Ponmariappan, S.;Kumar, Om;Singh, Lokendra
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.719-727
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    • 2011
  • Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

Effect of culture-promoting ingredients (CPI-107) on the culture of Mycobacterium tuberculosis (결핵균 배양에 대한 배양촉진물질(CPI-107)의 효과)

  • Seung Cheol Kim;Sezim Monoldorova;Bo-Young Jeon
    • Korean Journal of Veterinary Service
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    • v.46 no.1
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    • pp.29-34
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    • 2023
  • Mycobacterium tuberculosis complex (M. tuberculosis complex) is a causative agent of contagious chronic disease in a wide range of mammalian hosts, mainly cattle, goat, pigs, wildlife, and humans. The definite diagnosis of tuberculosis is made based on culture of M. tuberculosis, but it takes a long time. In the present study, we analyzed whether the detection time of M. tuberculosis could be reduced when cultured in the medium containing the culture-promoting ingredients-107 (CPI-107) using the BacT/Alert 3D system, an automatic culture system. The time to detection (TTD) tended to decrease as the added concentration of CPI-107 increase. In the case of low numbers of M. tuberculosis, it decreased by 21.0% at 1.2 mg/mL of CPI-107 and by 15.9% in the case of high numbers of M. tuberculosis. In the culture using clinically isolated M. tuberculosis strains, the shortening of the culture time by CPI was more evident. In conclusion, the detection time of M. tuberculosis was shortened in the medium added with CPI-107, and this could be used for isolation, culture and drug susceptibility test of M. tuberculosis.

Detection of Gene Interactions based on Syntactic Relations (구문관계에 기반한 유전자 상호작용 인식)

  • Kim, Mi-Young
    • The KIPS Transactions:PartB
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    • v.14B no.5
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    • pp.383-390
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    • 2007
  • Interactions between proteins and genes are often considered essential in the description of biomolecular phenomena and networks of interactions are considered as an entre for a Systems Biology approach. Recently, many works try to extract information by analyzing biomolecular text using natural language processing technology. Previous researches insist that linguistic information is useful to improve the performance in detecting gene interactions. However, previous systems do not show reasonable performance because of low recall. To improve recall without sacrificing precision, this paper proposes a new method for detection of gene interactions based on syntactic relations. Without biomolecular knowledge, our method shows reasonable performance using only small size of training data. Using the format of LLL05(ICML05 Workshop on Learning Language in Logic) data we detect the agent gene and its target gene that interact with each other. In the 1st phase, we detect encapsulation types for each agent and target candidate. In the 2nd phase, we construct verb lists that indicate the interaction information between two genes. In the last phase, to detect which of two genes is an agent or a target, we learn direction information. In the experimental results using LLL05 data, our proposed method showed F-measure of 88% for training data, and 70.4% for test data. This performance significantly outperformed previous methods. We also describe the contribution rate of each phase to the performance, and demonstrate that the first phase contributes to the improvement of recall and the second and last phases contribute to the improvement of precision.

Permanent Mycoplasma Removal Removel from Tissue Culture Cells: A Genetic Approach

  • Motr, Gabriele;Preininger, Alexandra;Himmelspach, Michele;Plaimauer, Barbara;Arbesser, Christine;York, Heinz;Dorner, Friedrich;Schlokat, Use
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.84-91
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    • 2000
  • Mycopasma contamination of tissue culture cells easily evades detection and, thus, represents a continous therat to cell biologists. In case where infected cell can not simply be replaced, attempts have to be made to eradicate mycoplacma from the tissue culture cells. A variety of anti-microbial agents have been shown to be toxic to mycoplasma strains ; however, cell associated mycoplasma are often protected from antibiotics at concentrations shown to be effective in vitro. Antibiotic concentrations high enough to be lethal to cell as sociated mycoplasmas frequently are also detrimentrations to the host cells, while moderately increased antibiotic levels tolerated by the host cells often lead to only temporary growth suppression and/or to the emergence of mycoplasma strains resistanct even to high concentrations of the antibiotis applied. Hare, a genetic approach for the elimination of mycoplasma from tissue culture cells that overcomes thens limitations is described. By expression of a selection marker conferring resistance to an otherwise toxic agent, Acholeplasma laidlawii infected BHK-21 cells used as the model system were enabled to temporarily tolerate antibiotic concentrations high enough to be lethal to cell associated mycopalsma while leaving the host cells unharmed. Upon successful mycoplasma eradicated, cultvation of the cured host cells in the absence of the selective agent yielded revertant cell clones that had regained susceptibillity to the toxic agent. Cressation of the selection marker expression was shown to result from the loss of the selection marker DNA, which is a consequence of the fact that the stable and permanent integration of foreign DNA in eucaryotic cell chrosomes is highly inefficient. Thus, the cells were cured from mycoplasma yet remained biochemically unaltered.

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An Agent System for Searching of Host Computer and Blocking Network Access in IPv6 Environment (IPv6 환경에서 호스트 탐색 및 네트워크 접속 차단 에이전트 시스템)

  • Chung, Youn-Ky;Moon, Hae-Eun
    • Journal of Korea Multimedia Society
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    • v.14 no.1
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    • pp.144-152
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    • 2011
  • As IPv4 addresses are exhausting, the use of IPv6 addresses is increasing. IPv6 environment provides address auto-configuration function. If addresses are allocated to each host automatically, network management system has difficulty in inspecting every IP of all devices and keeping the relevant informations. Also, as IP addresses are configured automatically, problems such as malicious users accessing network devices with no restriction can occur. To solve these problems, managing and blocking of malicious user is necessary. In this paper, we suggest agent system for searching of host computer and blocking network access which manages and protects the major network resources efficiently by searching host and blocking unauthorized host access to network in IPv6 environment. According to the test results of function of this agent system in IPv6 environment, we have checked that this system performs searching and blocking function normally.

Synthesis and Characterization of Novel pH-Sensitive Hydrogels Containing Ibuprofen Pen dents for Colon-Specific Drug Delivery

  • Mahkam, Mehrdad;Poorgholy, Nahid;Vakhshouri, Laleh
    • Macromolecular Research
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    • v.17 no.9
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    • pp.709-713
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    • 2009
  • The aim of this study was to develop novel intestinal specific drug delivery systems with pH sensitive swelling and drug release properties. The carboxyl group of ibuprofen was converted to a vinyl ester group by reacting ibuprofen and vinyl acetate as an acylating agent in the presence of catalyst. The glucose-6-acrylate-1, 2, 3, 4-tetraacetate (GATA) monomer was prepared under mild conditions. Cubane-1, 4-dicarboxylic acid (CDA) linked to two 2-hydroxyethyl methacrylate (HEMA) group was used as the crosslinking agent (CA). Methacrylic-type polymeric prodrugs were synthesized by the free radical copolymerization of methacrylic acid, vinyl ester derivative of ibuprofen (VIP) and GATA in the presence of cubane cross linking agent. The structure of VIP was characterized and confirmed by FTIR, $^1H$ NMR and $^{13}C$ NMR spectroscopy. The composition of the cross-linked three-dimensional polymers was determined by FTIR spectroscopy. The hydrolysis of drug polymer conjugates was carried out in cel-lophane membrane dialysis bags, and the in vitro release profiles were established separately in enzyme-free simulated gastric and intestinal fluids (SGF, pH 1 and SIF, pH 7.4). The detection of a hydrolysis solution by UV spectroscopy at selected intervals showed that the drug can be released by hydrolysis of the ester bond between the drug and polymer backbone at a low rate. Drug release studies showed that increasing the MAA content in the copolymer enhances the rate of hydrolysis in SIP. These results suggest that these polymeric prodrugs can be useful for the release of ibuprofen in controlled release systems.

An Agent based Emergency Warning System for Dealing With Defensive Information Warfare in Strategic Simulation Exercises (전략시뮬레이션 훈련에서의 방어적 정보전을 위한 에이전트 기반 위기경보시스템의 개발)

  • Lee Yong-Han;Kumara Soundar R.T.
    • Journal of Intelligence and Information Systems
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    • v.10 no.3
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    • pp.11-26
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    • 2004
  • Software for analyzing documents on the net to detect specific categories of occurrences is in great demand. In the current world where detecting terrorist threats is critical there is a great need for such systems. One of the critical application areas of such software is the automatic detection of a national infrastructure emergency. In this research an agent-based generic architecture for emergency warning systems is proposed and implemented. This system, called the National Infrastructure Emergency Warning System (NIEWS), is designed to analyze given documents, to detect threats, and to report possible threats with the necessary information to the appropriate users autonomously. In addition, a systematic analysis framework to detect emergencies on the subject of defensive information warfare is designated and implemented through a knowledge base. The developed system along with the knowledge base is implemented and successfully deployed to Strategic Crisis Exercise (SCE) at the United State Army War College (USAWC), saving a good amount of money by replacing human SMEs (subject matter experts) in the SCE.

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Analysis and Design of IP Traceback for Intrusion Response (침입대응을 위한 IP 역추적 시스템 분석 및 설계)

  • 이성현;이원구;이재광
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2004.05b
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    • pp.412-415
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    • 2004
  • As computers and networks become popular, corporation or country organization composes security network including various kinds information protection system to protect informations and resources from internet and is operating system and network. But current firewall and IDS(Intrusion Detection System) of the network level suffers from many vulnerabilities in internal computing informations and resources. In this paper, we design of ICMP-based Traceback System using a ICMP Traceback Message for efficiently traceback without change structure of routers. ICMP-based Traceback System. Create of ICMP message is managed by “Traceback Agent” mirroring port for router. Victim's systems that are received the message store it and “Traceback Manager” is detect a attack(like a DDoS). Using a information of this message starting a traceback and detecting a source of attacker, so response a attack.

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Detection of the Causal Agent of Bacterial Wilt, Ralstonia solanacearum in the Seeds of Solanaceae by PCR (가지과 종자에서 Ralstonia solanacearum의 검출을 위한 PCR 방법)

  • Cho, Jung-Hee;Yim, Kyu-Ock;Lee, Hyok-In;Baeg, Ji-Hyun;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.184-190
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    • 2011
  • Ralstonia solanacearum, a causal agent of bacterium wilt is very difficult to control once the disease becomes endemic. Thus, Ralstonia solanacearum is a plant quarantine bacterium in many countries including Korea. In this study, we developed PCR assays, which can detect Ralstonia solanacearum from the Solanaceae seeds. Primers RS-JH-F and RS-JH-R amplified specifically a 401 bp fragment only from Ralstonia solanacearum race 1 and race 3. The nested PCR primers, RS-JH-F-ne and RS-JH-R-ne that were designed inside of 1st PCR amplicon amplified specifically a 131 bp fragment only from Ralstonia solanacearum race 1 and race 3. The primers did not amplify any non-specific DNA from the seed extracts of the Solanaceae including tomato and pepper. When detection sensitivity were compared using the Solanaceae seeds inoculated with target bacteria artificially, the nested PCR method developed in this study 100 times more sensitive than ELISA and selective medium. Therefore, we believe that the PCR assays developed in this work is very useful to detect Ralstonia solanacearum in the Solanaceae seeds.