• 제목/요약/키워드: microplate

검색결과 151건 처리시간 0.023초

Microplate hybridization assay for detection of isoniazid resistance in Mycobacterium tuberculosis

  • Han, Hye-Eun;Lee, In-Soo;Hwang, Joo-Hwan;Bang, Hye-Eun;Kim, Yeun;Cho, Sang-Nae;Kim, Tae-Ue;Lee, Hye-Young
    • BMB Reports
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    • 제42권2호
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    • pp.81-85
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    • 2009
  • Early and accurate detection of drug resistant Mycobacterium tuberculosis can improve both the treatment outcome and public health control of tuberculosis. A number of molecular-based techniques have been developed including ones using probe molecules that target drug resistance-related mutations. Although these techniques are highly specific and sensitive, mixed signals can be obtained when the drug resistant isolates are mixed with drug susceptible isolates. In order to overcome this problem, we developed a new drug susceptibility test (DST) for one of the most effective anti-tuberculosis drug, isoniazid. This technique employed a microplate hybridization assay that quantified signals from each probe molecule, and was evaluated using clinical isolates. The evaluation analysis clearly showed that the microplate hybridization assay was an accurate and rapid method that overcame the limitations of DST based on conventional molecular techniques.

마이크로플레이트 기반 생물반응기 시스템 (MABOOMSTM)을 이용한 대장균 배양공정에서 용존산소, pH 및 세포농도의 온라인 모니터링 연구 (Study on Online Monitoring of Dissolved Oxygen, pH and Cell Concentration in E. coli Cultivation Processes Using MABOOMSTM)

  • 손옥재;이종일
    • KSBB Journal
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    • 제28권1호
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    • pp.24-30
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    • 2013
  • Dissolved oxygen, pH and cell concentration have been online monitored in cultivation processes with Escherichia coli by using a $MABOOMS^{TM}$ (microplate-based bioreactor with optical online monitoring systems). Fluorescent sensing membranes containing Ru ${(dpp)_3}^{2+}$ or HPTS were prepared with GA sol-gel matrix and coated into a well of a 24-well microplate. Fluorescence intensity was measured and correlated to the dissolved oxygen or pH. Cell concentrations were also online monitored by measuring optical reflectance at 650 nm. A well of a 24-well microplate could also be divided into 4 parts, each of which was coated with fluorescent sensing membranes for the detection of dissolved oxygen or pH. The 24-well microplate coated with fluorescent sensing membranes or a 4-divided sensing membrane. was used to online monitor the dissolved oxygen, pH and cell concentration during E. coli cultivations. The online monitoring results showed the characteristics of cell growth in cultivation processes very well.

Nonlinear bending of multilayer functionally graded graphene-reinforced skew microplates under mechanical and thermal loads using FSDT and MCST: A study in large deformation

  • J. Jenabi;A.R. Nezamabadi;M. Karami Khorramabadi
    • Structural Engineering and Mechanics
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    • 제90권3호
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    • pp.219-232
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    • 2024
  • In current study, for the first time, Nonlinear Bending of a skew microplate made of a laminated composite strengthened with graphene nanosheets is investigated. A mixture of mechanical and thermal stresses is applied to the plate, and the reaction is analyzed using the First Shear Deformation Theory (FSDT). Since different percentages of graphene sheets are included in the multilayer structure of the composite, the characteristics of the composite are functionally graded throughout its thickness. Halpin-Tsai models are used to characterize mechanical qualities, whereas Schapery models are used to characterize thermal properties. The microplate's non-linear strain is first calculated by calculating the plate shear deformation and using the Green-Lagrange tensor and von Karman assumptions. Then the elements of the Couple and Cauchy stress tensors using the Modified Coupled Stress Theory (MCST) are derived. Next, using the Hamilton Principle, the microplate's governing equations and associated boundary conditions are calculated. The nonlinear differential equations are linearized by utilizing auxiliary variables in the nonlinear solution by applying the Frechet approach. The linearized equations are rectified via an iterative loop to precisely solve the problem. For this, the Differential Quadrature Method (DQM) is utilized, and the outcomes are shown for the basic support boundary condition. To ascertain the maximum values of microplate deflection for a range of circumstances-such as skew angles, volume fractions, configurations, temperatures, and length scales-a parametric analysis is carried out. To shed light on how the microplate behaves in these various circumstances, the resulting results are analyzed.

생약 추출물에 의한 superoxide와 hydroxyl 라디칼 소거능 검색 방법의 개선 (An Improved method in Screening of Superoxide and Hydroxyl Radical Scavenging Activities of Plant Medicinal Extracts)

  • 이호섭;강대길
    • 생약학회지
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    • 제32권3호통권126호
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    • pp.253-256
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    • 2001
  • The present study was designed for the improvement of routine measurement of superoxide and hydroxyl radical scavenging activities utilized by a microplate reader. Superoxide radical scavenging activity by the ascorbic acid, which is a well-known superoxide scavenger, was determined in a dose-dependent manner. Hydroxyl radical scavenging activity by the thiourea, which is a well-known hydroxyl radical scavenger, was also well detected in a dose-dependent manner. Our results suggest that the use of microplate reader to assay the superoxide and hydroxyl radical scavenging activities improves the accuracy of data and enables the use of much smaller amounts of samples and/or reagents, with much simpler experimental procedure. Therefore, These methods appear to be suitable for screening of superoxide and hydroxyl radical scavenging activities in both the plant medicinal extracts and the isolated compounds.

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혈청 Progesterone 측정을 위한 효소면역분석법 개발에 관한 연구 (Studies on Development of Microplate-EIA for the Determination of Serum Progesterone)

  • 김정우;이욱연
    • 한국가축번식학회지
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    • 제17권4호
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    • pp.347-356
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    • 1994
  • A simpled and sensitive microplate enzyme immunoassay(EIA) was developed for the determination of progesterone concentration in serum, based on progesterone monoclonal antibody as anti-progesterone, horseradish peroxidase(HRP) as enzyme-label and tetramethylbenzidine(TMB) as substrate. The assay has a sensitivity of 5 pg-120pg/well and intra- and inter-assay coefficients of variation for progesterone standard curve (1.0ng~10.0ng/ml) were ranged 2.5~9.9% and 1.7.8.0%, respectively, determination coefficient of the regressio equation of our standard curve(R2=0.990$\pm$0.007) were high, and this is the same level as that of commercial kit(Hormonost Bio-Lab, Germany, R2=0.98~0.99). The progesterone concentration of serum determined by both kits (Work & Bio-Lab) were significantly correlated (r=0.95, P<0.01) although a little higher value were resulted in our kit than that of commercial kit. It generally is these results indicated that the microplate-EIA can be cused for the determination of progesterone in serum, as well as, for the determination of the early pregnancy diagnosis.

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Temperature-dependent nonlocal nonlinear buckling analysis of functionally graded SWCNT-reinforced microplates embedded in an orthotropic elastomeric medium

  • Barzoki, Ali Akbar Mosallaie;Loghman, Abbas;Arani, Ali Ghorbanpour
    • Structural Engineering and Mechanics
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    • 제53권3호
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    • pp.497-517
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    • 2015
  • In this study, nonlocal nonlinear buckling analysis of embedded polymeric temperature-dependent microplates resting on an elastic matrix as orthotropic temperature-dependent elastomeric medium is investigated. The microplate is reinforced by single-walled carbon nanotubes (SWCNTs) in which the equivalent material properties nanocomposite are estimated based on the rule of mixture. For the carbon-nanotube reinforced composite (CNTRC) plate, both cases of uniform distribution (UD) and functionally graded (FG) distribution patterns of SWCNT reinforcements are considered. The small size effects of microplate are considered based on Eringen's nonlocal theory. Based on orthotropic Mindlin plate theory along with von K$\acute{a}$rm$\acute{a}$n geometric nonlinearity and Hamilton's principle, the governing equations are derived. Generalized differential quadrature method (GDQM) is applied for obtaining the buckling load of system. The effects of different parameters such as nonlocal parameters, volume fractions of SWCNTs, distribution type of SWCNTs in polymer, elastomeric medium, aspect ratio, boundary condition, orientation of foundation orthtotropy direction and temperature are considered on the nonlinear buckling of the microplate. Results indicate that CNT distribution close to top and bottom are more efficient than those distributed nearby the mid-plane for increasing the buckling load.

Development of PCR-microplate Hybridization Assay for Detection of Mycobacterium tuberculosis

  • Lee, In-Soo;Cho, Een-Jin;Cho, Sang-Nae;Kim, Tae-Ue;Lee, Hye-Young
    • 대한의생명과학회지
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    • 제15권4호
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    • pp.295-300
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    • 2009
  • Tuberculosis caused by Mycobacterium tuberculosis (MTB) still remains to be the most dreadful infectious disease affecting almost every country. In the present study, we developed a simple and rapid but accurate and sensitive assay method for detecting MTB using microplate hybridization assay. For this, a selective region of the rpoB gene was used to design PCR primers, and MTB and Mycobacterium genus-specific probe molecules. The specificity of the assay was confirmed using fifteen different mycobacterial reference strains and twelve different non-mycobacterial reference strains, and the sensitivity was determined to be 100 fg using genomic DNA of MTB reference strain, H37Rv. Subsequently, a total of 62 sputum samples with diverse smear scores and culture positive results were used to evaluate the kit performance. In brief, the specificity and the sensitivity of the assay were 100% and 98.4%, respectively.

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Microplate법에 의한 된장유래의 항혈전 펩타이드 탐색 (Screening of Antithrombotic Peptides from Soybean Paste by the Microplate Method)

  • 손동화;이경애;김승호;안창원;남희섭;이형재;신재익
    • 한국식품과학회지
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    • 제28권4호
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    • pp.684-688
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    • 1996
  • 전통식품인 된장으로부터 혈소판 응집 저해활성을 갖는 항혈전 펩타이드를 탐색하고자 하였다. 증류수로 추출한 된장액을 한외 여과하여 얻은 분자량 3,000 dalton 이하의 여액에 대하여, SD흰쥐의 세척혈소판 (washed platelet)을 이용하여 ADP 자극으로 유도되는 혈소판응집의 저해정도를 aggregometer에서의 탁도로 측정하였을 때, $96\;{\mu}g/ml$로 처리시 90% 가량의 응집 저해효과가 확인되었다. 추출물을 Dowex 50W X-2 강양이온 교환칼럼을 이용하여 pyridine-formic acid 완충액의 pH (2.8-5.0) 및 몰농도(0.1-3.0M)를 단계별로 증가시키면서 용출시켜 19개의 획분(No.B-18)으로 분리하였다. 이들 획분의 활성을 microplate를 이용한 방법으로 측정하였을 때, 전체적으로 앞의 방법보다는 감도가 낮게 나타났으나 모든 분획에서 응집억제 효과가 확인되었다. 이때 용량반응 곡선으로부터 구한 각 획분의 $IC_{50}$$10-1,000\;{\mu}g/ml)$의 범위를 보였으며, 대부분의 획분들은 양성대조구인 RGDS $(IC_{50},\;205\;{\mu}g/ml)$보다 높은 활성을 나타내었다. 또한, 후반부의 염기성 획분$(IC_{50},\;10-20\;{\mu}g/ml)$에서 된장추출물$(IC_{50},\;30\;{\mu}g/ml)$보다 활성이 높게 나타났다. 이들 중 활성이 가장 높은 16번 획분을 아미노산 분석한 결과, 펩타이드 부분에 histidine, arginine, alanine 등이 높은 비율로 존재하였다.

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고추의 카로티노이드 함량 분석을 위한 신속·간편 마이크로플레이트법 (A Simple and Fast Microplate Method for Analysis of Carotenoids Content in Chili Pepper (Capsicum annuum L.))

  • 홍순철;한정헌;이준대;안율균;양은영;채수영;김수;윤재복
    • 원예과학기술지
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    • 제31권6호
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    • pp.807-812
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    • 2013
  • 본 연구에서는 폴리스틸렌 재질의 96-well 마이크로플레이트와 ELISA 판독기를 이용하여 고추의 카로티노이드 함량을 간편하게 분석할 수 있는 방법을 개발하고자 하였다. 분광광도계를 이용하지 않고 450nm 필터가 장착된 ELISA 판독기로도 카로티노이드 분석이 가능하였으나, 카로티노이드 추출 시 사용하는 아세톤으로 인해 마이크로플레이트의 표면이 불투명하게 변색되었다. 그러나 아세톤 색소추출물을 메탄올로 10배 희석한 용액을 사용할 경우, 플레이트 표면과 용액 내 카로티노이드 성분의 변화 없이 카로티노이드 측정이 가능하였고, ASTA-20.1법으로 측정된 결과와도 높은 상관관계를 보였다. 한편, 카로티노이드 추출 조건으로는 고춧가루 0.1g을 아세톤 10mL에 첨가하고 실온에서 4시간 동안 교반하는 것이 적합하였다. 본 실험의 ELISA 판독기를 이용한 색소분석법(Microplate법으로 명명)은 대량의 시료를 처리할 수 있을 뿐만 아니라 아세톤 폐액의 발생량을 ASTA-20.1법의 1/10 수준으로 줄일 수 있어 실제 고추 육종에서 고색소 계통을 선발할 때 매우 유용할 것으로 판단된다.

Nonlinear stability of non-axisymmetric functionally graded reinforced nano composite microplates

  • Loghman, Abbas;Arani, Ali Ghorbanpour;Barzoki, Ali Akbar Mosallaie
    • Computers and Concrete
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    • 제19권6호
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    • pp.677-687
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    • 2017
  • The nonlinear buckling response of nano composite anti-symmetric functionally graded polymeric microplate reinforced by single-walled carbon nanotubes (SWCNTs) rested on orthotropic elastomeric foundation with temperature dependent properties is investigated. For the carbon-nanotube reinforced composite (CNTRC) microplate, a uniform distribution (UD) and four types of functionally graded (FG) distribution are considered. Based on orthotropic Mindlin plate theory, von Karman geometric nonlinearity and Hamilton's principle, the governing equations are derived. Generalized differential quadrature method (GDQM) is employed to calculate the non-linear buckling response of the plate. Effects of FG distribution type, elastomeric foundation, aspect ratio (thickness to width ratio), boundary condition, orientation of foundation orthotropy and temperature are considered. The results are validated. It is found that the critical buckling load without elastic medium is significantly lower than considering Winkler and Pasternak medium.