• Title/Summary/Keyword: method detection limit

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Multiplex PCR Assay for Simultaneous Detection of Korean Quarantine Phytoplasmas

  • Kim, Young-Hwan;Win, Nang Kyu;Back, Chang-Gi;Yea, Mi-Chi;Yim, Kyu-Ock;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.367-371
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    • 2011
  • Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/${\mu}l$). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.

A New Analytical Method to Determine the Purity of Synthetic Fluorophores using Single Molecule Detection Technique

  • Song, Nam-Yoong;Kim, Hyong-Ha;Park, Tae-Sook;Yoon, Min-Joong
    • Journal of Photoscience
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    • v.12 no.2
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    • pp.87-93
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    • 2005
  • A new assay technique to distinguish between pure compounds and the isomeric mixtures has been suggested using single molecule (SM) fluorescence detection technique. Since the number of emission spots in a fluorophorespread film prepared from a genuine dye solution was determined by experimental condition, the deviation of spot numbers from the expected values could be considered to be an indication of lower purity of the sample solution. The lower limit of sample concentration for this assay was determined to be $5{\times}10^{-10}$ M to show uniform number of expected spots within 10% uncertainties in our experimental condition. An individual fluorescence intensity distribution for a mixture of isomers having doubly different emissivities was simulated by adding distributions obtained from Cy3 and nile red (NR) independently. The result indicated that the mixture could be identified from the pure compounds through the difference in the number of Gaussian functions to fit the distribution. This new assay technique can be applied to the purity test for synthetic biofluorophores which are usually prepared in small quantities not enough for classical ensemble assays.

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Quantitative Detection of Salmonella typhimurium Contamination in Milk, Using Real-Time PCR

  • JUNG SUNG JE;KIM HYUN-JOONG;KIM HAE-YEONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1353-1358
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    • 2005
  • A rapid and quantitative real-time PCR was developed to target the invasion A (invA) gene of Salmonella spp. We developed quantitative standard curves based on plasmids containing the invA gene. Based on these curves, we detected Salmonella spp. in artificially contaminated buffered peptone water (BPW) and milk samples. We were able to determine the invA gene copy number per ml of food samples, with the minimum detection limit of $4.1{\times}10^{3}$ copies/ml of BPW and $3.3{\times}10^{3}$ copies/ml of milk. When applied directly to detect and quantify Salmonella spp. in BPW and milk, the present real-time PCR assay was as sensitive as the plate count method; however, copy numbers were one to two logs higher than the colony-forming units obtained by the plate count methods. In the present work, the real-time PCR assay was shown to significantly reduce the total time necessary for the detection of Salmonella spp. in foods and to provide an important model for other foodborne pathogens.

Direct Detection of Shigella flexneri and Salmonella typhimurium in Human Feces by Real-Time PCR

  • Yang, Young-Geun;Song, Man-Ki;Park, Su-Jeong;Kim, Suhng-Wook
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1616-1621
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    • 2007
  • We have established a SYBR Green-based realtime PCR method using AnyDirect solution, which enhances PCR from whole blood, for direct amplification of the virA gene of Shigella flexneri and the invA gene of Salmonella typhimurium from human feces without prior DNA purification. When we compared the efficiency of conventional or realtime PCR amplification of the virA and invA genes from the supernatant of boiled feces supplemented with S. flexneri and S. typhimurium in the presence or absence of AnyDirect solution, amplification products were detected only in reactions to which AnyDirect solution had been added. The detection limit of real-time PCR was $1{\times}10^4\;CFU/g$ feces for S. flexneri and $2{\times}10^4\;CFU/g$ feces for S. typhimurium; this sensitivity level was comparable to other studies. Our real-time PCR assay with AnyDirect solution is simple, rapid, sensitive, and specific, and allows simultaneous detection of S. flexneri and S. typhimurium directly from fecal samples without prior DNA purification.

A Development of Video Monitoring System on Real Time (실시간 영상감시 시스템 개발)

  • Cho, Hyun-Seob
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.8 no.2
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    • pp.240-244
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    • 2007
  • Non-intrusive methods based on active remote IR illumination fur eye tracking is important for many applications of vision-based man-machine interaction. One problem that has plagued those methods is their sensitivity to lighting condition change. This tends to significantly limit their scope of application. In this paper, we present a new real-time eye detection and tracking methodology that works under variable and realistic lighting conditions. Based on combining the bright-pupil effect resulted from IR light and the conventional appearance-based object recognition technique, our method can robustly track eyes when the pupils are not very bright due to significant external illumination interferences. The appearance model is incorporated in both eyes detection and tracking via the use of support vector machine and the mean shift tracking. Additional improvement is achieved from modifying the image acquisition apparatus including the illuminator and the camera.

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Determination of Biotin by HPLC (고성능 액체크로마토크래피를 이용한 Biotin의 정량)

  • Kim, Dong-Soo;Lee, Young-Ja;Jeong, Dong-Youn;Lee, Dong-Yup;Ahn, Moon-Kyu
    • Analytical Science and Technology
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    • v.16 no.6
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    • pp.499-503
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    • 2003
  • A high performance liquid chromatography gradient elution method with fluorescence detection for the determination of biotin in pure form and pharmaceutical preparations has been developed. BrMDMC gives intense fluorescence and the fluorescence was monitored with excitation at 360 nm and emission at 410 nm. The calibration curve for biotin shows good linearity over the range of 5 ~ 400 ng with correlation coefficient of 0.999. The detection limit of biotin was 2 ng and the result of recovery was 98.75% with relative standard deviation of 1.1%.

Detection of Sequence-Specific Gene by Multi-Channel Electrochemical DNA Chips

  • Zhang, Xuzhi;Ji, Xinming;Cui, Zhengguo;Yang, Bing;Huang, Jie
    • Bulletin of the Korean Chemical Society
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    • v.33 no.1
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    • pp.69-75
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    • 2012
  • Five-channel electrochemical chips were fabricated based on the Micro-electromechanical System (MEMS) technology and were used as platforms to develop DNA arrays. Different kinds of thiolated DNA strands, whose sequences were related to white spot syndrome virus (WSSV) gene, were separately immobilized onto different working electrodes to fabricate a combinatorial biosensor system. As a result, different kinds of target DNA could be analyzed on one chip via a simultaneous recognition process using potassium ferricyanide as an indicator. To perform quantitative target DNA detection, a limit of 70 nM (S/N=3) was found in the presence of 600 nM coexisting noncomplementary ssDNA. The real samples of loop-mediated isothermal amplification (LAMP) products were detected by the proposed method with satisfactory result, suggesting that the multichannel chips had the potential for a high effective microdevice to recognize specific gene sequence for pointof-care applications.

Examination of ivermectin residues in raw milk after skin administration (원유중 Ivermectin 구충제의 잔류실태 조사)

  • Bark, Jun-Jo;Youk, Ji-Hea;Kim, Hu-Kyoung;Park, Hye-Won;Kim, In-Kyung;Lee, Woo-Seong
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.421-428
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    • 2007
  • This study was conducted two kinds of aims: 1) to modify the analytical methods (conditions) by high performance liquid chromatography - fluorescence detector for the detection of residual ivermectin in raw milk, 2) to provide basic information for the evaluation of standard of the residual ivermectin in raw milk. It could be considerable that negative ion spectra can be better method in the LC/MS analysis for the detection of residues, Characteristic daughter ions were observed in negative ion spectra, however, linear line was not formed in positive ion one. Three Holstein cows ($500{\pm}10kg$) were applied to commercial ointment of ivermectin just one time at the first day of test, and residues in raw milk were examined for 20day after administration. The limit of detection (LOD) was 0.65ng (n=5) by HPLC/FLD, and recovery rates were $87.85%{\sim}99.47%$. The peak was observed at the 4th day, and residues lasted to the end. Thus ivermectin was prohibited when lactating.

Determination of Nebramycin Factor 2,4,5,5',6 and Kanamycin A in Fermentation Broth of Streptoalloteichus hindustanus ATCC 31218 Mutant Using 2,4-Dinitrofluorobenzene(DNFB) as a Derivatizing Agent by High Performance Liquid Chromatography (HPLC법에 의한 2,4-디니트로플루오로벤젠을 유도체화제로 한 Streptoalloteichus hindustanus ATCC 31218 변이균의 배양액 중 네브라마이신 펙터 2,4,5,5',6, 가나마이신 A 분석)

  • 박영근;박명용;김승철;양호길
    • YAKHAK HOEJI
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    • v.37 no.1
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    • pp.1-8
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    • 1993
  • A procedure for the high-performance liquid chromatographic determination of Nebramycin factors in fermentation broth of Streptoalloteichus hindustanus ATCC 31218 mutant was investigated using pre-column derivatization and LTV detection. The method is based on pre-column derivatization of Nebramycin factors with 2,4-dinitrofluorobenzene(DNFB) in the presence of Tris (hydroxymethyl)aminoethane. The chromatographic separation of derivatives of Nebramycin factors and unknown impurities is achieved using reversed-phase column (NOVA-PAK $C_{18}$, Waters Co.) and AcCN : H$_{2}$O : AcOH (53.0:46.5:0.5) as a mobile phase. The mixture of these derivatives were separated within 35 minutes and the optimum wavelength($\lambda_{max}$ ) of the UV detector was 353 nm. The linearity of response for derivatives of Nebramycin factors is demonstrated for concentrations up to 500 $\mu\textrm{g}$/ml and the relative standard deviation is less than 0.79%. Detection limit was 1.67 ng for the 10 $\mu\textrm{l}$ sample volume employed.

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Automatic Pancreas Detection on Abdominal CT Images using Intensity Normalization and Faster R-CNN (복부 CT 영상에서 밝기값 정규화 및 Faster R-CNN을 이용한 자동 췌장 검출)

  • Choi, Si-Eun;Lee, Seong-Eun;Hong, Helen
    • Journal of Korea Multimedia Society
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    • v.24 no.3
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    • pp.396-405
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    • 2021
  • In surgery to remove pancreatic cancer, it is important to figure out the shape of a patient's pancreas. However, previous studies have a limit to detect a pancreas automatically in abdominal CT images, because the pancreas varies in shape, size and location by patient. Therefore, in this paper, we propose a method of learning various shapes of pancreas according to the patients and adjacent slices using Faster R-CNN based on Inception V2, and automatically detecting the pancreas from abdominal CT images. Model training and testing were performed using the NIH Pancreas-CT Dataset, and intensity normalization was applied to all data to improve pancreatic detection accuracy. Additionally, according to the shape of the pancreas, the test dataset was classified into top, middle, and bottom slices to evaluate the model's performance on each data. The results show that the top data's mAP@.50IoU achieved 91.7% and the bottom data's mAP@.50IoU achieved 95.4%, and the highest performance was the middle data's mAP@.50IoU, 98.5%. Thus, we have confirmed that the model can accurately detect the pancreas in CT images.