• Title/Summary/Keyword: Conjugate Method

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Screening of zearalenone-producing strains by ELISA method (ELISA법에 의한 zearalenone 생성균주의 검색)

  • Kim, Sung-Young;Chung, Sun-Hee;Chung, Duck-Hwa
    • Applied Biological Chemistry
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    • v.36 no.1
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    • pp.7-10
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    • 1993
  • ELISA method was applied for the screening of zearalenone producing strains. The developed ELISA was as follow: $125\;{\mu}l$ of diluted solution (1 : 500) of antibody was added to each microtiter well and incubated overnight at $40^{\circ}C$. For direct competitive ELISA, samples and zearalenone-peroxidase conjugate were mixed in a 1 : 1 ratio, and a $100\;{\mu}l$ of aliquot was then added to antisera-coated wells. Plates were incubated for 30 minutes at $37^{\circ}C$, and wells washed 6 times, and $100\;{\mu}l$ of ABTS substrates was added. Plates were incubated for antother 15 minutes at $37^{\circ}C$, and $100\;{\mu}l$ of stopping reagent was added to the wells and absorbance was recorded at 410nm on ELISA Reader. Among 19 strains showed zearalenone-producing ability by ELISA, 3 strains (R-5, C-46, S-134) produced more than 50 ng/ml of zearalenone.

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Effect of Cross Phase Modulation on Channel Compensation in 320 Gbps Intensity Modulation / Direct Detection WDM Transmission Systems (320 Gbps 강도 변조 직접 검파 WDM 시스템의 채널 보상에서 상호 위상 변조의 영향)

  • 이성렬;김지웅;손성찬
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.8 no.5
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    • pp.1134-1140
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    • 2004
  • In this paper, we investigated the effect of cross phase modulation(XPM) on compensation for WDM channel distortion due to chromatic dispersion, self phase modulation and XPM as a function of fiber dispersion coefficient and modulation format in 320 Gbps WDM systems. The considered WDM transmission system is based on mid-span spectral inversion(MSSI) compensation method, which has highly nonlinear dispersion shifted fiber(HNL-DSF) as nonlinear medium of optical phase conjugator (OPC) in the mid-way of total transmission line. We confirmed that the maximum channel input power resulting 1 dB eye opening penalty is reduced due to XPM effect on channel distortion, even if MSSI method was applied to WDM system. But, we confirmed that the effect of XPM on channel distortion becomes decrease as fiber dispersion coefficient of WDM system becomes larger. Futhermore, we confirmed that NRZ is better than RZ as a modulation format for similarly compensating overall WDM channels in WDM system with large fiber dispersion coefficient in order to minimize the effect of the XPM on channel distortion.

Establishment of Neospora agglutination test for serologic diagnosis of neosporosis (Neospora응집반응을 이용한 네오스포라증의 혈청학적 진단)

  • Kang, Min-Soo;Kim, Jae-Hoon;Hwang, Woo-Suk;Nam, Ho-Woo;Youn, Hee-Jeong;Bae, Jong-Hee;Kim, Dae-Yong
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.677-681
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    • 2003
  • Currently, both the indirect fluorescent antibody test (IFAT) and enzyme linked immunosorbent assay (ELISA) have been used to detect Neospora caninum antibodies. Several factors such as the buffers, the conjugate, the pattern of fluorescence, and the cross reactivity with other apicornplexan protozoan, may result in poorly correlated data. The present study was undertaken to develop and evaluate the Neospora agglutination test (NAT) for the detection and quantification of IgG antibodies to N. caninum from various animal species. Compared to the ELISA method, the NAT with a cutoff value of 1:512 gave a high index of coincidence (kappa=0.807) and no cross reactivity to Toxoplasma gondii antiserum. Hence, this NAT method, which did not require a species-specific secondary antibody and expensive tools, would be easily available for the detection of antibodies to N. caninllm of various animal species.

The Method of Reducing Echo Time in 3D Time-of-flight Angiography

  • Park, Sung-Hong;Park, Jung-Il;Lee, Heung-Kyu
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2002.09a
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    • pp.367-369
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    • 2002
  • We have designed ramp profile excitation pulse based on the Shinnar-Le Roux (SLR) algorithm. The algorithm provides many advantages to pulse designers. The first advantage is the freedom of deciding the amplitudes, frequencies, and ripple sizes of stopband, passband, and transition band of pulse profile. The second advantage is the freedom of deciding the pulse phase, more specifically, minimum phase, linear phase, maximum phase, and any phase between them. The minimum phase pulse is the best choice in the case of 3D TOF, because it minimizes the echo time, which implies the best image quality in the same MR examination condition. In addition, the half echo technique is slightly modified in our case. In general, using the half echo technique means that the acquired data size is half and the rest part can be filled with complex conjugate of acquired data. But in our case, the echo center is just shifted to left, which implies the reduction of echo time, and the acquired data size is the same as the one without using the half echo technique. In this case, the increase of right part of data leads to improvement of the resolution and the decrease of left part of data leads to decrease of signal to noise ratio. Since in the case of 3D TOF, the signal to noise ratio is sufficiently high and the resolution is more important than signal to noise ratio, the proposed method appears to be significantly affective and gives rise to the improved high resolution angiograms.

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A Study on the Stability and Mechanism of Three-Hinge Failure (Three-Hinge 파괴의 메커니즘 및 안정성에 관한 분석)

  • Moon, Joon-Shik;Park, Woo-Jeong
    • Journal of the Korean Geotechnical Society
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    • v.33 no.4
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    • pp.5-15
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    • 2017
  • Three-hinge failure occurs in a jointed rock slope with a joint set parallel with slope and a conjugate joint set. Limit Equilibrium Method (LEM) and Finite Element Method (FEM) which are commonly used for slope design, are not suitable for evaluating stability against three-hinge failure, and this study performed parametric study to analyze the failure mechanism and to find influence factors causing three-hinge failure using UDEC which is a commercial two-dimensional DEM based numerical program. Numerical analyses were performed for various joint structural conditions and joint properties as well as ground water conditions. It was found that pore water pressure is the main factor triggering the three-hinge failure and the mode of failure depends on friction angle of basal joint and bedding joint set. The results obtained from this study can be used for adequate and economic footwall slope reinforcement design and construction.

A STUDY ON A MULTI-LEVEL SUBSTRUCTURING METHOD FOR COMPUTATIONS OF FLUID FLOW (유동계산을 위한 다단계 부분 구조법에 대한 연구)

  • Kim J.W.
    • Journal of computational fluids engineering
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    • v.10 no.2
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    • pp.38-47
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    • 2005
  • Substructuring methods are often used in finite element structural analyses. In this study a multi-level substructuring(MLSS) algorithm is developed and proposed as a possible candidate for finite element fluid solvers. The present algorithm consists of four stages such as a gathering, a condensing, a solving and a scattering stage. At each level, a predetermined number of elements are gathered and condensed to form an element of higher level. At the highest level, each sub-domain consists of only one super-element. Thus, the inversion process of a stiffness matrix associated with internal degrees of freedom of each sub-domain has been replaced by a sequential static condensation of gathered element matrices. The global algebraic system arising from the assembly of each sub-domain matrices is solved using a well-known iterative solver such as the conjugare gradient(CG) or the conjugate gradient squared(CGS) method. A time comparison with CG has been performed on a 2-D Poisson problem. With one domain the computing time by MLSS is comparable with that by CG up to about 260,000 d.o.f. For 263,169 d.o.f using 8 x 8 sub-domains, the time by MLSS is reduced to a value less than $30\%$ of that by CG. The lid-driven cavity problem has been solved for Re = 3200 using the element interpolation degree(Deg.) up to cubic. in this case, preconditioning techniques usually accompanied by iterative solvers are not needed. Finite element formulation for the incompressible flow has been stabilized by a modified residual procedure proposed by Ilinca et al.[9].

Fluorometric Detection of Low-Abundance EGFR Exon 19 Deletion Mutation Using Tandem Gene Amplification

  • Kim, Dong-Min;Zhang, Shichen;Kim, Minhee;Kim, Dong-Eun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.662-667
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    • 2020
  • Epidermal growth factor receptor (EGFR) mutations are not only genetic markers for diagnosis but also biomarkers of clinical-response against tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). Among the EGFR mutations, the in-frame deletion mutation in EGFR exon 19 kinase domain (EGFR exon 19-del) is the most frequent mutation, accounting for about 45% of EGFR mutations in NSCLCs. Development of sensitive method for detecting the EGFR mutation is highly required to make a better screening for drug-response in the treatment of NSCLC patients. Here, we developed a fluorometric tandem gene amplification assay for sensitive detection of low-abundance EGFR exon 19-del mutant genomic DNA. The method consists of pre-amplification with PCR, thermal cycling of ligation by Taq ligase, and subsequent rolling circle amplification (RCA). PCR-amplified DNA from genomic DNA samples was used as splint DNA to conjugate both ends of linear padlock DNA, generating circular padlock DNA template for RCA. Long stretches of ssDNA harboring multiple copies of G-quadruplex structure was generated in RCA and detected by thioflavin T (ThT) fluorescence, which is specifically intercalated into the G-quadruplex, emitting strong fluorescence. Sensitivity of tandem gene amplification assay for detection of the EGFR exon 19-del from gDNA was as low as 3.6 pg, and mutant gDNA present in the pooled normal plasma was readily detected as low as 1% fraction. Hence, fluorometric detection of low-abundance EGFR exon 19 deletion mutation using tandem gene amplification may be applicable to clinical diagnosis of NSCLC patients with appropriate TKI treatment.

Development of an Enzyme-Linked Immunosorbent Assay for the Iletection of Aflatoxin $B_1$ (Aflatoxin $B_1$의 검출을 위한 효소면역측정법의 개발)

  • 손동화;박애란;서병철;김진철;이인원;남영중;허우덕
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.225-232
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    • 1992
  • In order to develop an enzyme-linked immunosorbent assay(ELISA) for detecting aflatoxin $B_1(AFB_1)$, we produced and purified antibodies, thereafter established and evaluated methods of direct and indirect competitive ELISA. Anti-AFB, antisera, produced by immunizing rabbits with $AFB_1$-1-(0-carboxymethy1)oxime-bovine serum albumin conjugate ($AFB_1$-BSA), were removed of anti-BSA antibodies by quantitative precipitation reaction and further purified by ammonium sulfate precipitation and DEAE-Sephadex A-50 ion exchange chromatography. Purified IgG fractions were used as anti-$AFB_1$ antibodies. The antibodies, whose titer was deterrnined extremely high above $2 \times 10^6$, showed low cross-reactivity of 3~34% against $AFB_1$ analogues such as G2, B2, and GI. From the standard curves of direct and indirect competitive ELISA for AFBI, the detection ranges were found 0.2~20 and 1~10, 000 ng/ml(ppb) respectively. In their sensitivity, stability, simplicity, and rapidity, the direct method was more suitable than the indirect method for practical use.

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Standardization of an enzyme-linked immunosorbent assay for detection of antibody to avian reticuloendotheliosis virus (세망내피증 바이러스 항체검출을 위한 ELISA 표준화)

  • Sung, Haan Woo;Lee, Su Jeong
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.569-574
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    • 2005
  • Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

Electroanalytical Measurement of TEDA (Triethylenediamine) in the Masks of War

  • Ariani, Zahra;Honarmand, Ebrahim;Mostaanzadeh, Hossein;Motaghedifard, Mohammadhassan;Behpour, Mohsen
    • Journal of Electrochemical Science and Technology
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    • v.8 no.1
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    • pp.43-52
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    • 2017
  • In this paper, for the first time, the electroanalytical study of Triethylenediamine, TEDA was done on a typically graphene modified carbon paste electrode (Gr/CPE) in pH=10.5 of phosphate buffer solutions (PBS). The surface morphology of the bare and modified electrodes was characterized by scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The electro-oxidation of TEDA was investigated at the surface of modified electrode. The results revealed that the oxidation peak current of TEDA at the surface of Gr/CPE is 2.70 times than that shown at bare-CPE. A linear calibration plot was observed in the range of 1.0 to 202.0 ppm. In this way, the detection limit was found to be 0.18 ppm. The method was then successfully applied to determination of TEDA in aqueous samples obtained from two kinds of activated carbon from the masks of war. On the other hand, density functional theory (DFT) method at B3LYP/6-311++G** level of theory and a conductor-like Polarizable Continuum Model (CPCM) was used to calculate the $pK_a$ values of TEDA. The energies of lowest unoccupied molecular orbital ($E_{LUMO}$) and highest occupied molecular orbital ($E_{HOMO}$), gap energy (${\Delta}E$) and some thermodynamic parameters such as Gibbs free energy of TEDA and its conjugate acid ($HT^+$) were calculated. The results of calculated $pK_a$ were found to be in good agreement with the experimental values.