• 제목/요약/키워드: cross-reactions

검색결과 288건 처리시간 0.031초

Genomics-based Sensitive and Specific Novel Primers for Simultaneous Detection of Burkholderia glumae and Burkholderia gladioli in Rice Seeds

  • Lee, Chaeyeong;Lee, Hyun-Hee;Mannaa, Mohamed;Kim, Namgyu;Park, Jungwook;Kim, Juyun;Seo, Young-Su
    • The Plant Pathology Journal
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    • 제34권6호
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    • pp.490-498
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    • 2018
  • Panicle blight and seed rot disease caused mainly by Burkholderia glumae and Burkholderia gladioli is threatening rice cultivation worldwide. The bacteria have been reported as seed-borne pathogens from rice. Accurate detection of both pathogens on the seeds is very important for limiting the disease dissemination. Novel primer pairs targeting specific molecular markers were developed for the robust detection of B. glumae and B. gladioli. The designed primers were specific in detecting the target species with no apparent cross-reactions with other related Burkholderia species at the expected product size. Both primer pairs displayed a high degree of sensitivity for detection of B. glumae and B. gladioli separately in monoplex PCR or simultaneously in duplex PCR from both extracted gDNA and directly preheated bacterial cell suspensions. Limit of detection was as low as 0.1 ng of gDNA of both species and $3.86{\times}10^2cells$ for B. glumae and $5.85{\times}10^2cells$ for B. gladioli. On inoculated rice seeds, the designed primers could separately or simultaneously detect B. glumae and B. gladioli with a detection limit as low as $1.86{\times}10^3cells$ per rice seed for B. glumae and $1.04{\times}10^4cells$ per rice seed of B. gladioli. The novel primers maybe valuable as a more sensitive, specific, and robust tool for the efficient simultaneous detection of B. glumae and B. gladioli on rice seeds, which is important in combating rice panicle blight and seed rot by early detection and confirmation of the dissemination of pathogen-free rice seeds.

전자충격반응을 포함하는 플라즈마 화학반응을 고려한 용량결합형 산소플라즈마의 전산모사 연구 (Simulation Study of Capacitively Coupled Oxygen Plasma with Plasma Chemistry including Detailed Electron Impact Reactions)

  • 김헌창
    • 공업화학
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    • 제22권6호
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    • pp.711-717
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    • 2011
  • 전자충격반응을 고려한 three moment 플라즈마 모델과 전기적 중성성분의 반응을 고려한 유체 유동 모델을 결합하여 용량결합형 산소플라즈마에 대한 2차원적 전산모사 연구를 수행하였다. 전자의 에너지에 의하여 좌우되는 전자충격반응에 대한 반응속도는 전자와 $O_2$ 및 O 사이의 전자충돌단면적으로부터 계산되었다. 플라즈마 모델과 유체 유동 모델을 결합하고 상세한 반응메커니즘을 포함시킴으로써 전하를 띠는 전자와 이온($O_2{^+}$, $O^+$, $O_2{^-}$, and $O^-$) 그리고 기저상태의 산소($O_2$ and O)뿐만 아니라 $O_2(a^1{\Delta}_g)$, $O_2(b^1{{\Sigma}_g}^+)$, $O(^1D)$, $O(^1S)$ 등과 같이 산소플라즈마 특성에 중요한 역할을 하는 준안정상태 성분들의 시공간적 분포를 예측할 수 있었다. 또한 산소플라즈마의 전산모사로부터 sheath 경계에 이중층이 존재함을 확인할 수 있었다.

A LAMP-SNP Assay Detecting C580Y Mutation in Pfkelch13 Gene from Clinically Dried Blood Spot Samples

  • Khammanee, Thunchanok;Sawangjaroen, Nongyao;Buncherd, Hansuk;Tun, Aung Win;Thanapongpichat, Supinya
    • 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.

Rapid Detection of Clostridium tetani by Recombinase Polymerase Amplification Using an Exo Probe

  • Guo, Mingjing;Feng, Pan;Zhang, Liqun;Feng, Chunfeng;Fu, Jie;Pu, Xiaoyun;Liu, Fei
    • Journal of Microbiology and Biotechnology
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    • 제32권1호
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    • pp.91-98
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    • 2022
  • Tetanus is a potentially fatal public health illness resulted from the neurotoxins generated by Clostridium tetani. C. tetani is not easily culturable and culturing the relevant bacteria from infected wounds has rarely been useful in diagnosis; PCR-based assays can only be conducted at highly sophisticated laboratories. Therefore, a real-time recombinase polymerase amplification assay (Exo-RPA) was constructed to identify the fragments of the neurotoxin gene of C. tetani. Primers and the exo probe targeting the conserved region were designed, and the resulting amplicons could be detected in less than 20 min, with a detection limit of 20 copies/reaction. The RPA assay displayed good selectivity, and there were no cross-reactions with other infectious bacteria common in penetrating wounds. Tests of target-spiked serum and pus extract revealed that RPA is robust to interfering factors and has great potential for further development for biological sample analysis. This method has been confirmed to be reliable for discriminating between toxic and nontoxic C. tetani strains. The RPA assay dramatically improves the diagnostic efficacy with simplified device architecture and is a promising alternative to real-time PCR for tetanus detection.

Establishment of multiplex RT-PCR for differentiation between rabies virus with and that without mutation at position 333 of glycoprotein

  • Yang, Dong-Kun;Kim, Ha-Hyun;Lee, Siu;Yoo, Jae-Young
    • Journal of Veterinary Science
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    • 제21권2호
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    • pp.22.1-22.9
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    • 2020
  • Rabid raccoon dogs (Nyctereutes procyonoides koreensis) have been responsible for animal rabies in South Korea since the 1990s. A recombinant rabies vaccine strain, designated as ERAGS, was constructed for use as a bait vaccine. Therefore, new means of differentiating ERAGS from other rabies virus (RABV) strains will be required in biological manufacturing and diagnostic service centers. In this study, we designed two specific primer sets for differentiation between ERAGS and other RABVs based on mutation in the RABV glycoprotein gene. Polymerase chain reaction analysis of the glycoprotein gene revealed two DNA bands of 383 bp and 583 bp in the ERAGS strain but a single DNA band of 383 bp in the field strains. The detection limits of multiplex reverse transcription polymerase chain reaction (RT-PCR) were 80 and 8 FAID50/reaction for the ERAGS and Evelyn-Rokitnicki-Abelseth strains, respectively. No cross-reactions were detected in the non-RABV reference viruses, including canine distemper virus, parvovirus, canine adenovirus type 1 and 2, and parainfluenza virus. The results of multiplex RT-PCR were 100% consistent with those of the fluorescent antibody test. Therefore, one-step multiplex RT-PCR is likely useful for differentiation between RABVs with and those without mutation at position 333 of the RABV glycoprotein gene.

Echinococcus granulosus Protoscolex DM9 Protein Shows High Potential for Serodiagnosis of Alveolar Echinococcosis

  • Kim, Jeong-Geun;Han, Xiumin;Kong, Yoon
    • Parasites, Hosts and Diseases
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    • 제60권1호
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    • pp.25-34
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    • 2022
  • Alveolar echinococcosis (AE) caused by infection with E. multilocularis metacestode, represents one of the most fatal helminthic diseases. AE is principally manifested with infiltrative, proliferating hepatic mass, resembling primary hepatocellular carcinoma. Sometimes metastatic lesions are found in nearby or remote tissue. AE diagnosis largely depends on imaging studies, but atypical findings of imaging features frequently require differential diagnosis from other hepatic lesions. Serological tests may provide further evidence, while obtaining reliable AE materials is not easy. In this study, alternative antigens, specific to AE were identified by analyzing E. granulosus protoscolex proteins. An immunoblot analysis of E. granulosus protoscolex showed that a group of low-molecular-weight proteins in the range from 14 kDa to 16 kDa exhibited a sensitive and specific immune response to AE patient sera. Partial purification and proteomic analysis indicated that this protein group contained myosin, tubulin polymerization promoting protein, fatty-acid binding protein, uncharacterized DM9, heat shock protein 90 cochaperone tebp P-23, and antigen S. When the serological applicability of recombinant forms of these proteins was assessed using enzyme-linked immunosorbent assay, DM9 protein (rEgDM9) showed 90.1% sensitivity (73/81 sera tested) and 94.5% specificity (172/181 sera tested), respectively. rEgDM9 showed weak cross-reactions with patient sera from the transitional and chronic stages of cystic echinococcosis (3 to 5 stages). rEgDM9 would serve as a useful alternative antigen for serodiagnosis of both early- and advanced-stage AE cases.

다당류 및 TGase를 활용한 동결 무지개송어육(Oncorhynchus mykiss)의 물성개선 및 저장성 향상 효과 (Effect of Texture Improvement and Shelf Life Extension of Frozen Rainbow Trout Oncorhynchus mykiss Treated with TGase and Polysaccharides)

  • 이종봉;박혜민;안병규;이우진;인정진;한형구;손승아;배연주;심길보
    • 한국수산과학회지
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    • 제56권4호
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    • pp.505-511
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    • 2023
  • This study investigated the effect of transglutaminase (TGase) and polysaccharide kappa carrageenan on the texture, chemical, and microbiological qualities of refrigerated unmarketable rainbow trout Oncorhynchus mykiss. Gel strength increased substantially in TGase-treated samples, and was adding kappa carrageenan resulted in no significant difference. SDS-PAGE results confirmed that the myosin heavy chain band with a molecular weight of 205-250 kDa was weakened in trout meat treated with 1% TGase, which led to cross-linking reactions between proteins. The volatile basic nitrogen (VBN) increased in all samples during storage at 4℃ for 10 days; however, the samples treated with 0.5% and 1% kappa carrageenan had the lowest VBN. The viable cell count increased in all samples treated with TGase and kappa carrageenan; however, an increase in TGase enzyme and kappa carrageenan concentration successfully hindered total bacteria growth. Thus, adding 1% TGase and 1% kappa carrageenan to refrigerated unmarketable rainbow trout formulations can optimize quality characteristics.

Positron Annihilation Spectroscopy of Active Galactic Nuclei

  • Doikov, Dmytry N.;Yushchenko, Alexander V.;Jeong, Yeuncheol
    • Journal of Astronomy and Space Sciences
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    • 제36권1호
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    • pp.21-33
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    • 2019
  • This paper focuses on the interpretation of radiation fluxes from active galactic nuclei. The advantage of positron annihilation spectroscopy over other methods of spectral diagnostics of active galactic nuclei (therefore AGN) is demonstrated. A relationship between regular and random components in both bolometric and spectral composition of fluxes of quanta and particles generated in AGN is found. We consider their diffuse component separately and also detect radiative feedback after the passage of high-velocity cosmic rays and hard quanta through gas-and-dust aggregates surrounding massive black holes in AGN. The motion of relativistic positrons and electrons in such complex systems produces secondary radiation throughout the whole investigated region of active galactic nuclei in form of cylinder with radius R= 400-1000 pc and height H=200-400 pc, thus causing their visible luminescence across all spectral bands. We obtain radiation and electron energy distribution functions depending on the spatial distribution of the investigated bulk of matter in AGN. Radiation luminescence of the non-central part of AGN is a response to the effects of particles and quanta falling from its center created by atoms, molecules and dust of its diffuse component. The cross-sections for the single-photon annihilation of positrons of different energies with atoms in these active galactic nuclei are determined. For the first time we use the data on the change in chemical composition due to spallation reactions induced by high-energy particles. We establish or define more accurately how the energies of the incident positron, emitted ${\gamma}-quantum$ and recoiling nucleus correlate with the atomic number and weight of the target nucleus. For light elements, we provide detailed tables of all indicated parameters. A new criterion is proposed, based on the use of the ratio of the fluxes of ${\gamma}-quanta$ formed in one- and two-photon annihilation of positrons in a diffuse medium. It is concluded that, as is the case in young supernova remnants, the two-photon annihilation tends to occur in solid-state grains as a result of active loss of kinetic energy of positrons due to ionisation down to thermal energy of free electrons. The single-photon annihilation of positrons manifests itself in the gas component of active galactic nuclei. Such annihilation occurs as interaction between positrons and K-shell electrons; hence, it is suitable for identification of the chemical state of substances comprising the gas component of the investigated media. Specific physical media producing high fluxes of positrons are discussed; it allowed a significant reduction in the number of reaction channels generating positrons. We estimate the brightness distribution in the ${\gamma}-ray$ spectra of the gas-and-dust media through which positron fluxes travel with the energy range similar to that recorded by the Payload for Antimatter Matter Exploration and Light-nuclei Astrophysics (PAMELA) research module. Based on the results of our calculations, we analyse the reasons for such a high power of positrons to penetrate through gas-and-dust aggregates. The energy loss of positrons by ionisation is compared to the production of secondary positrons by high-energy cosmic rays in order to determine the depth of their penetration into gas-and-dust aggregations clustered in active galactic nuclei. The relationship between the energy of ${\gamma}-quanta$ emitted upon the single-photon annihilation and the energy of incident electrons is established. The obtained cross sections for positron interactions with bound electrons of the diffuse component of the non-central, peripheral AGN regions allowed us to obtain new spectroscopic characteristics of the atoms involved in single-photon annihilation.

망간산화물(Birnessite)을 이용한 1- Indanone 제거 시 페놀계 반응매개체와 휴믹산(HA) 영향 평가 (Effect of Phenolic Mediators and Humic Acid on the Removal of 1-Indanone Using Manganese Oxide)

  • 최찬규;엄원숙;신현상
    • 대한환경공학회지
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    • 제34권7호
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    • pp.445-453
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    • 2012
  • 본 연구에서는 PAHs 오염토양의 자연 풍화 및 화학적 생물학적 처리과정에서 반응부산물로 흔히 발견되는 PAH-케톤화합물인 1-indanon (1-ID)을 대상으로 페놀계 반응매개체 존재 하에서의 망간산화물에 의한 산화 변환 제거특성 및 용존 자연유기물인 휴믹산(HA)의 존재에 따른 영향을 조사하였다. 반응성 평가 실험은 수용액 상에서 회분식(10 mg/L 1-ID, 0.3 mM phenolic mediators, $1.0g/L\;{\delta}-MnO_2$, at pH 5)으로 수행 하였으며, 페놀계의 반응매개체(phenolic mediator)는 자연산 페놀화합물로서 휴믹물질의 모델 화합물로서도 널리 사용되고 있는 11종을 사용하였다. 실험결과 1-ID은 망간산화물 자체에 대하여는 비반응성을 띠었으나 페놀계 반응매개체 존재 하에서 교차-결합(cross-coupling)반응을 통해 제거됨을 HPLC 분석을 통해 확인하였으며, 1-ID의 제거율은 반응 2일 경과 후 9.2~71.2%범위에서 페놀계 반응매개체의 구조적 특성에 따라 다르게 나타났다. 각 반응매개체 존재 하에서의 1-ID의 교차결합 반응은 유사1차 반응 속도식을 따랐으며, 초기 반응속도 상수 값($K_{int}$, $hr^{-1}$)은 0.48~15.0의 넓은 범위에서 나타났다. 1-ID의 제거효율(제거율, 속도상수)은 -OH, $-OCH_3$ 등 전자주게(electron donating) 작용기를 포함하는 반응매개체에서 높았으며, -COOH, -CHO 등 전자받게(electron withdrowing) 작용기를 포함하는 반응매개체일수록 낮았다. 또한 동일 반응 조건에서 HA 존재에 따른 영향을 검토한 결과 낮은 HA 농도(< 2 mg/L) 조건에서는 1-ID 제거효율의 상승효과를 보였으나 전체적으로는 HA 주입 농도가 증가할수록 교차 결합 반응효율이 저하됨을 확인하였다.

전극 표면에 부착된 IPN 형태의 전해질 고분자의 제조 및 그들의 감습특성 (Preparation of IPN-type Polyelectrolyte Films Attached to the Electrode Surface and Their Humidity-Sensitive Properties)

  • 한대상;공명선
    • 폴리머
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    • 제34권6호
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    • pp.565-573
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    • 2010
  • IPN 구조를 가지는 감습성 고분자 전해질로 사용하기 위하여 디브로모알칸과 가교가 가능한 copoly(2-(dimethylamino) ethyl methacrylate)(DAEMA)/butyl acrylate(BA)와 광가교가 가능한 copoly(methyl methacrylate) (MMA)/BA/2-(cinnamoyloxy)ethyl methacryate(CEMA)를 제조하였다. 전극의 기재 표면에 광조사에 의한 IPN-감습성 전해질의 부착을 위하여 3-(triethoxysilyl)propyl cinnamate(TESPC)을 전극표면에 처리하였다. IPN 구조의 감습성 고분자 필름은 copoly(DAEMA/BA), copoly(MMA/BA/CEMA) 및 1,4-dibromobutane(DBB) 가교제의 혼합 용액에 침적한 후에 UV 조사와 동시에 가열하여 제조하였다. IPN-전해질 고분자의 전극 기재와의 부착은 광화학적 $[2{\pi}+2{\pi}]$ 환화반응에 의하여 진행하였다. 얻어진 습도센서는 20~95%RH의 영역에서 매우 높은 감도와 1.5%RH 이하의 작은 히스테리시스를 보여주었다. 또한, 33~94%RH 사이에서 가습과 제습과정의 응답 및 회복 속도는 각각 48초와 65초로 나타나 매우 빠름을 알 수 있었다. 그 밖에 감습액 중의 공중합체의 농도, 가교제의 양, 가교반응 시간 등이 내수성을 포함한 감습성질에 미치는 영향을 조사하였다.