• 제목/요약/키워드: Molecular Detection

검색결과 1,108건 처리시간 0.034초

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.

무균성 뇌막염과 뇌염으로 입원한 성인 환자 뇌척수액에서 중합효소 연쇄반응에 의한 HSV, VZV, HHV-6의 검출 (Detection of Herpes Simplex Virus, Varicella-Zoster Virus and Human Herpes Virus-6 by PCR in Cerebrospinal Fluid from Hospitalized Adult Patients with Aseptic Meningitis or Encephalitis)

  • 박혜경;우소연;김현진;정영해
    • 대한바이러스학회지
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    • 제30권3호
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    • pp.171-178
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    • 2000
  • Herpes simplex virus, Varicella zoster virus and Human herpes virus-6 caused central nervous system infections and latent infections but there is no data of the 3 viruses being tested from the same cerebrospinal fluid samples with aseptic meningitis or encephalitis in adults patients. These viruses produced similar neurologic symptoms but difficulties existed in differentiating of etiologic agents and therefore the viruses needed to be detected in the early state. Herpes simplex virus encephalitis (HSVE) in adults, if not treated promptly was fatal. If treated with antiviral drugs in the early phase of encephalitis, neurologic sequales decreased by 65%. Recently, a PCR method for detection of HSVE with CSF was developed. VZV primary and secondary infections caused neurologic symptoms of encephalitis or meningitis. The second frequency of adult encephalitis that caused VZV were reported. HHV-6 caused CNS latent infection that was studied with normal adults brains. But there is no data of HSV, VZV and HHV-6 for aseptic meningitis and encephalitis of Korean adults through etiologic study. We cultured CSFs on HEp-2 cells and simultaneously tested for HSV PCR, VZV nested PCR and HHV-6 PCR with 8 specific primers. The PCR results of CSF from meningitis Korean adults were 13/19 (68.4%) for HSV, 10/19 (52.6%) for VZV and 12/19 (63.2%) for HHV-67/19 (36.8%) cases were triple infected HSV PCR, VZV PCR and HHV-6 PCR positive; 3/19 (15.8%) cases were dual infected HSV PCR and HHV-6 PCR positive; 1119 (0.5%) cases was VZV PCR positive. Strong viral DNA amplification of CSF means a causative virus may be present in aseptic meningitis or encephalitis patients and may cause clinical neurologic symptoms. HSV and HHV-6 viruses detection rate were higher than VZV by PCR with CSFs.

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Development of a Rapid Automated Fluorescent Lateral Flow Immunoassay to Detect Hepatitis B Surface Antigen (HBsAg), Antibody to HBsAg, and Antibody to Hepatitis C

  • Ryu, Ji Hyeong;Kwon, Minsuk;Moon, Joung-Dae;Hwang, Min-Woong;Lee, Jeong-Min;Park, Ki-Hyun;Yun, So Jeong;Bae, Hyun Jin;Choi, Aeran;Lee, Hyeyoung;Jung, Bongsu;Jeong, Juhee;Han, Kyungja;Kim, Yonggoo;Oh, Eun-Jee
    • Annals of Laboratory Medicine
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    • 제38권6호
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    • pp.578-584
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    • 2018
  • Background: Accurate, rapid, and cost-effective screening tests for hepatitis B virus (HBV) and hepatitis C virus (HCV) infection may be useful in laboratories that cannot afford automated chemiluminescent immunoassays (CLIAs). We evaluated the diagnostic performance of a novel rapid automated fluorescent lateral flow immunoassay (LFIA). Methods: A fluorescent LFIA using a small bench-top fluorescence reader, Automated Fluorescent Immunoassay System (AFIAS; Boditech Med Inc., Chuncheon, Korea), was developed for qualitative detection of hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs), and antibody to HCV (anti-HCV) within 20 minutes. We compared the diagnostic performance of AFIAS with that of automated CLIAs-Elecsys (Roche Diagnostics GmbH, Penzberg, Germany) and ARCHITECT (Abbott Laboratories, Abbott Park, IL, USA)-using 20 seroconversion panels and 3,500 clinical serum samples. Results: Evaluation with the seroconversion panels demonstrated that AFIAS had adequate sensitivity for HBsAg and anti-HCV detection. From the clinical samples, AFIAS sensitivity and specificity were 99.8% and 99.3% for the HBsAg test, 100.0% and 100.0% for the anti-HBs test, and 98.8% and 99.1% for the anti-HCV test, respectively. Its agreement rates with the Elecsys HBsAg, anti-HBs, and anti-HCV detection assays were 99.4%, 100.0%, and 99.0%, respectively. AFIAS detected all samples with HBsAg genotypes A-F and H and anti-HCV genotypes 1, 1a, 1b, 2a, 2b, 4, and 6. Cross-reactivity with other infections was not observed. Conclusions: The AFIAS HBsAg, anti-HBs, and anti-HCV tests demonstrated diagnostic performance equivalent to current automated CLIAs. AFIAS could be used for a large-scale HBV or HCV screening in low-resource laboratories or low-to middle-income areas.

Prenatal Population Screening for Fragile X Carrier and the Prevalence of Premutation Carriers in, Korea

  • Han, Sung-Hee;Heo, Yun-Ah;Yang, Young-Ho;Kim, Young-Jin;Cho, Han-Ik;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • 제9권2호
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    • pp.73-77
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    • 2012
  • Purpose: Fragile X carrier detection before or at early pregnancy through a wide screening program may not only confer a risk of having offspring with Fragile X syndrome (FXS), but may also confer a risk for Fragile X-associated primary ovarian insufficiency and Fragile X-associated tremor/ataxia syndrome. However, prior to the implementation of such a program, the carrier prevalence in a population and the availability of effective screening test should be evaluated. The aim of our study was to determine the prevalence of premutation carriers and to evaluate the feasibility of screening test. Materials and Methods: The blood samples were obtained from 8,641 pregnant women with no family history of mental retardation. We performed a three-primer CGG repeat primed (RP) PCR using the AmplideX$^{TM}$ FMR1 PCR kit (Asuragen, Inc. Austin, TX, USA). Samples showing full mutation alleles were reflexed to Southern blot analysis for methylation status and sizing. Results: Among the 8,641 women, we found 8 premutation carriers (1:1,090, 0.09%) and 46 women with an intermediate allele (1:190, 0.53%). No woman was found to carry the fully mutated allele. All the detected alleles were within the CGG repeat range of 8-117. Among the 8,641 samples, 29 and 30 CGG repeats represent 66.6% of all cases. The CGG RP PCR method provides robust detection of expanded alleles and resolves allele zygosity, thus minimizing the number of samples that require Southern blot analysis. Conclusion: This is the first study that has focused on the prevalence of FXS premutation carriers and FMR1 allele distribution in normal pregnant women. These data have important implications for population-based fragile X carrier screening in Korea.

종 특이 DNA probe를 이용한 버섯 세균성 갈반병 병원균(Pseudomonas tolaasii)의 검출 (Detection of Pseudomonas tolaasii Causing Brown Blotch Disease of Mushroom with Species-specific DNA Probe)

  • 권순우;고승주;전명숙;강희완;오세종;장후봉;류진창
    • 한국균학회지
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    • 제27권2호통권89호
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    • pp.132-137
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    • 1999
  • 본 연구는 느타리버섯 갈반병 병원균인 pseudomonas tolaasii의 진단을 위한 분자 marker를 개발하기 위해 수행되었다. 세균의 반복염기서열과 펙틴 분해효소 유전자로부터 제작된 여러개의 primer들을 이용해 식용버섯으로부터 분리된 Pseudomonas종들로부터 DNA 다형성을 유도한 바펙틴 분해 효소로부터 제작된 PEU1 primer는 다른 Pseudomonas종들로부터 P. tolaasii를 구분시키는 다형성 밴드를 생성하였다. P. tolaasii 6균주에 공통적으로 나타나는 1.0kb와 0.4kb의 두 가지 밴드를 pGEM-T에 클로닝하고 이들을 각기 pPTOP1과 pPTOP2로 명명하고 probe로 이용하였다. 0.4kb 크기의 pPTOP2의 삽입 DNA는 P. tolaasii 6균주와 hybridization이 이루어진 반면 다른 Pseudomonas종과는 반응하지 않았다. 0.4kb크기의 pPTOP2의 삽입 DNA를 probe로 이용해 dot blot hybridization한 결과 P. tolaasii는 $1.5{\times}10^3\;cfu$까지 검출이 가능함을 확인하였다.

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Change of Molecular Weight of Organic Matters through Unit Water Treatment Process and Associated Chlorination Byproducts Formation

  • Sohn, Jin-Sik;Kang, Hyo-Soon;Han, Ji-Hee;Yoon, Yeo-Min
    • Environmental Engineering Research
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    • 제12권5호
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    • pp.224-230
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    • 2007
  • The objectives of this study were to evaluate the change of molecular weight (MW) profiles in natural organic matter (NOM) through various treatment processes (coagulation, granular activated carbon (GAC), and ozonation) using high performance size exclusion chromatography based on ultraviolet absorbance and dissolved organic detection (HPSEC-UVA-DOC). In addition, relationships between MW profiles and disinfection by-production (DBP) formation were evaluated. Each treatment process results in significant different effects on NOM profiles. Coagulation is effective to remove high molecular weight NOM, while GAC is effective to remove low molecular weight NOM. Ozonation removes only a small portion of NOM, while it induces a significant reduction of UV absorbance due to breakdown of the aromatic groups. All treated waters are chlorinated, and chlorination DBPs such as trihalomethanes (THMs) and haloacetic acids (HAAs) are measured under formation potential conditions. Both THM and HAA formation potentials were significantly reduced through the coagulation process. GAC was more effective to reduce THM formation compared to HAA formation reduction, while ozonation showed significant HAA reduction compared to THM reduction.

MAP kinase kinase kinase as a positive defense regulator in rice-blast fungus interactions

  • Kim, Jung-A;Jung, Young-Ho;Lee, Joo-Hee;Jwa, Nam-Soo
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2004년도 The 2004 KSPP Annual Meeting & International Symposium
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    • pp.48-52
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    • 2004
  • We have found the role of rice mitogen-activated protein kinase kinase kinase (MAPKKK), OsEDR1, as controling hypersensitive response (HR) and increased disease resistance to rice blast fungus Magnaporthe grisea. Generation of transgenic rice plants through introduction of the over-expression construct of OsEDR1 using Agrobacterium-mediated transformation results in lesion mimic phenotype. Up-regulation of defense mechanism was detected through detection of increased transcription level of rice PBZ1 and PR1a. Inoculation of rice blast fungus on the lesion mimic transgenic lines displayed significantly increased resistance. The disease symptoms were arrested like HR responses which are commonly detected in the incompatible interactions. High accumulation of phenolic compounds around developing lesions was detected under UV light. There was variation among transgenic lines on the timing of lesion progression as well as the lesion numbers on the rice leaves. Transgenic lines with few lesions also show increased resistance as well as equal amount of grain yields compared to that of wild type rice cultivar Nipponbare. This is the first report of the MAPKKK as a positive regulator molecule on defense mechanism through inducing HR-like cell death lesion mimic phenotype. The application of OsEDR1 is highly expected for the development of resistant cultivars against rice pathogens.

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DLPC 인지질 단분자막의 변위전류 특성 연구 (I) (A Study on Displacement Current Characteristics of DLPC Monolayer (I))

  • 송진원;이경섭;최용성
    • 전기학회논문지
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    • 제56권1호
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    • pp.117-122
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    • 2007
  • LB method is one of the most interesting technique to arrange certain molecular groups at precise position relative to others. Also, the LB deposition technique can fabricate extremely thin organic films with a high degree of control over their thickness and molecular architecture. In this way, new thin film materials can be built up at the molecular level, and the relationship between these artificial structures and the properties of materials can be explored. In this paper, evaluation of physical properties was made for dielectric relaxation phenomena by the detection of the surface pressures and displacements current on the monolayer films of phospolipid monomolecular DLPC. Lipid thin films were manufacture by detecting deposition for the accumulation and the current was measured after the electric bias was applied across the manufactured MIM device. It is found that the phospolipid monolayer of dielectric relaxation takes a little time and depend on the molecular area. When electric bias is applied across the manufactured MIM device by the deposition condition of phospolipid mono-layer, it wasn't breakdown when the higher electric field to impress by increase of deposition layers.

Comparison of Two Methods to Extract DNA from Formalin-Fixed, Paraffin-Embedded Tissues and their Impact on EGFR Mutation Detection in Non-small Cell Lung Carcinoma

  • Hu, Yu-Chang;Zhang, Qian;Huang, Yan-Hua;Liu, Yu-Fei;Chen, Hong-Lei
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권6호
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    • pp.2733-2737
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    • 2014
  • Objective: Molecular pathology tests are often carried for clinicopathological diagnosis and pathologists have established large collections of formalin-fixed, paraffin-embedded tissue (FFPE) banks. However, extraction of DNA from FFPE is a laborious and challenging for researchers in clinical laboratories. The aim of this study was to compare two widely used DNA extraction methods: using a QIAamp DNA FFPE kit from Qiagen and a Cobas Sample Preparation Kit from Roche, and evaluated the effect of the DNA quality on molecular diagnostics. Methods: DNA from FFPE non-small cell lung carcinoma tissues including biopsy and surgical specimens was extracted with both QIAamp DNA FFPE and Cobas Sample Preparation Kits and EGFR mutations of non-small cell lung carcinomas were detected by real-time quantitative PCR using the extracted DNA. Results and Conclusion: Our results showed that DNA extracted by QIAamp and Cobas methods were both suitable to detect downstream EGFR mutation in surgical specimens. Howover, Cobas method could yield more DNA from biopsy specimens, and gain much better EGFR mutation results.

Simultaneous Determination of Alkoxyalcohols in Wet Wipes Using Static Headspace Gas Chromatography and Mass Spectrometry

  • Lee, Soojin;Pyo, Heesoo;Chung, Bong Chul;Kim, Haidong;Lee, Jeongae
    • Bulletin of the Korean Chemical Society
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    • 제35권11호
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    • pp.3280-3288
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    • 2014
  • Alkoxyalcohols are used as solvents or preservatives in various consumer products such as wet wipes. The metabolites of alkoxyalcohols are known to be chronically toxic and carcinogenic to animals. Thus, an analytical method is needed to monitor alkoxyalcohols in wet wipes. The aim of this study was to develop a simultaneous analytical method for 14 alkoxyalcohols using headspace gas chromatography coupled with mass spectrometry to analyze the wet wipes. This method was developed by comparing with various headspace extraction parameters. The linear calibration curves were obtained for the method ($r^2$ > 0.995). The limit of detection of alkoxyalcohols ranged from 2 to $200ng\;mL^{-1}$. The precision of the determinative method was less than 18.20% coefficient of variation both intra and inter days. The accuracy of the method ranged from 82.86% to 119.83%. (2-Methoxymethylethoxy)propanol, 2-phenoxyethanol, and 1-phenoxy-2-propanol were mainly detected in wet wipes.