• Title/Summary/Keyword: Real samples

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Rapid Detection Method for Human Rotavirus from Vegetables by a Combination of Filtration and Integrated Cell Culture/Real-Time Reverse Transcription PCR (Filtration과 Integrated Cell Culture/Real-Time Reverse Transcription PCR 기법을 이용한 채소류에서 Human Rotavirus 신속 검출)

  • Hyeon, Ji-Yeon;Chon, Jung-Whan;Song, Kwang-Young;Hwang, In-Gyun;Kwak, Hyo-Sun;Lee, Jung-Soo;Kim, Moo-Sang;Lee, Jung-Bok;Seo, Kun-Ho
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.117-123
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    • 2011
  • The purpose of this study was to evaluate and compare different elution and concentration methods for optimization of human rotavirus (HRV) detection method using real-time RT-PCR and cell culture techniques. The leafy vegetable samples (lettuce, Chinese cabbage) were artificially inoculated with HRV. Viruses were extracted from the vegetables by two different elution buffers, buffer A (100 mM Tris-HCl, 50 mM glycine, 3% beef extract, pH 9.5) and buffer B (250 mM Threonine, 300 mM NaCl, pH 9.5), and the extracted viruses were concentrated by filtration and PEG precipitation sequentially. To determine infectivity of the viruses, the viruses recovered from the samples were infected to the MA-104 cells, and integrated cell culture real-time RT-PCR was performed at 1, 48, 72, 96, 120, 144, 168 h post-infection (p.i.). The elution buffer A was more efficient in extracting the virus from the produce samples tested than the buffer B, 29.54% and 18.32% of recoveries, respectively. The sensitivity of real-time RT-PCR method was markedly improved when the virus was concentrated by the filtration method. When the viruses were eluted and concentrated by buffer A and filtration, respectively, the average recovery rate was approximately 51.89%. When the viruses recovered from samples were infected to MA-104 cell, infectious HRV was detected within 48 h p.i. by ICC/real-time RT-PCR, whereas cytopathic effects were not observed until 72 h p.i. The optimized detection method evaluated in this study could be useful for rapid and reliable detection of HRV in fresh produce products and applied for detection of other food-borne viruses.

Comparison of Standard Culture Method and Real-time PCR Assay for Detection of Staphylococcus aureus in Processed and Unprocessed Foods (가공식품과 비가공식품에서의 황색포도상구균 검출을 위한 배지법과 Real-time PCR법의 비교)

  • Lee, Jae-Hoon;Song, Kwang-Young;Hyeon, Ji-Yeon;Hwang, In-Gyun;Kwak, Hyo-Sun;Han, Jeong-A;Chung, Yun-Hee;Seo, Kun-Ho
    • Food Science of Animal Resources
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    • v.30 no.3
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    • pp.410-418
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    • 2010
  • Staphylococcus aureus is one of the major pathogens that can cause staphylococcal infection and food poisoning. In this study, we compared conventional culture methods and real-time PCR for detection of S. aureus in artificially inoculated milk, sausage, raw pork, and vegetable salad. The performance of a coagulase test for confirming S. aureus was also compared with a colony PCR test. Bulk food samples (500 g each) were artificially inoculated with S. aureus and divided into 20 samples (25 g or mL each). All samples were added to tryptic soy broth (225 mL/sample) with 10% NaCl and incubated at $37^{\circ}C$ for 24 h. After the enrichment, broth cultures were streaked onto Baird-Parker (BP) agar with egg yolk tellulite, and incubated at $37^{\circ}C$ for 24 h. In addition, 1 mL of broth cultures was collected to perform real-time PCR. Two suspicious colonies from the BP agar were picked up and plated on nutrient agar and incubated at $37^{\circ}C$ for 24 h followed, by a coagulase confirmation test and a colony PCR analysis. There were no statistical differences between culture methods and realtime PCR in food samples with low background microflora, such as milk and sausage. However, a significant statistical difference was found between the culture methods and real-time PCR for raw pork and vegetable salad. Furthermore, the colony PCR test of the presumptive colonies on BP agar for confirming S. aureus is more accurate and efficient than the coagulase test for unprocessed foods.

Anodization of Aluminium Samples in Boric Acid Solutions by Optical Interferometry Techniques

  • Habib, K.
    • Corrosion Science and Technology
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    • v.4 no.6
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    • pp.217-221
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    • 2005
  • In the present investigation, holographic interferometry was utilized for the first time to monitor in situ the thickness of the oxide film of aluminium samples during anodization processes in boric acid solutions. The anodization process (oxidation) of the aluminium samples was carried out by the technique of the electrochemical impedance spectroscopy(EIS), in different concentrations of boric acid (0.5-5.0% $H_3BO_3$) at room temperature. In the mean time, the real-time holographic interferometry was used to measure the thickness of anodized (oxide) film of the aluminium samples in solutions. Consequently, holographic interferometry is found very useful for surface finish industries especially for monitoring the early stage of anodization processes of metals, in which the thickness of the anodized film of the aluminium samples can be determined without any physical contact. In addition, measurements of electrochemical values such as the alternating current (A.C) impedance(Z), the double layer capacitance($C_{dl}$), and the polarization resistance(Rp) of anodized films of aluminium samples in boric acid solutions were made by the electrochemical impedance spectroscopy(EIS). Attempts to measure electrochemical values of Z, Cdl, and Rp were not possible by holographic interferometry in boric acid especially in low concentrations of the acid. This is because of the high rate of evolutions of interferometric fringes during the anodization process of the aluminium samples in boric acid, which made measurements of Z, Cdl, and Rp are difficult.

Comparison of Loop-mediated Isothermal Amplification and Korea Standard Food Codex (KFSC) Method for Detection of Salmonella Typhimurium, Listeria monocytogenes Artificially Inoculated in Yuk-hwe and Yuk-sashimi (육회와 육사시미에 접종된 Salmonella Typhimurium와 Listeria monocytogenes 검출을 위한 Loop-mediated isothermal amplification와 식품공전의 배지 시험법, real-time PCR의 검출 성능 비교)

  • Gwak, Seung-Hae;Lee, So-Young;Kim, Jin-Hee;Oh, Se-Wook
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.277-282
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    • 2019
  • The object of this study is to compare the performance of the 3M Molecular Detection Assay 2 (3M MDA 2) and the Korea Standard Food Codex (KSFC) Method (i.e., isolation media and real-time PCR) in detecting Salmonella Typhimurium and Listeria monocytogenes in traditional Korean foods. Yuk-hwe and Yuk-sashimi (types of raw beef dishes) were artificially inoculated with $10^0-10^4CFU/25g$ of L. monocytogenes and S. Typhimurium. Citrobacter freundii and Listeria innocua were used as competitive microflora. After enrichment, the samples were analyzed using 3M MDA 2 and real-time PCR. All samples inoculated at concentrations of $10^0-10^4CFU/25g$ without competitive microflora were positive for S. Typhimurium and L. monocytogenes, as detected by 3M MDA 2 and Korea Standard Food Codex (KFSC) Method. In addition, part of the samples were positive for the presence of C. freundii and L. innocua. The 3M MDA 2 - Salmonella and Korea Standard Food Codex (KFSC) Method showed similar detection performances in Yuk-hwe and Yuk-sashimi. The 3M MDA 2 method for Salmonella and Listeria, which is a LAMP-based technology, can be used for rapid detection of S. Typhimurium and L. monocytogenes in raw beef. LAMP bioluminescence assays provide results on the subsequent day and are simple to use compared with the Korea Standard Food Codex (KFSC) Method, particularly in terms of DNA preparation.

On Quantifies Estimation Using Ranked Samples with Some Applications

  • Samawi, Hani-M.
    • Journal of the Korean Statistical Society
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    • v.30 no.4
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    • pp.667-678
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    • 2001
  • The asymptotic behavior and distribution for quantiles estimators using ranked samples are introduced. Applications of quantiles estimation on finding the normal ranges (2.5% and 97.5% percentiles) and the median of some medical characteristics and on finding the Hodges-Lehmann estimate are discussed. The conclusion of this study is, whenever perfect ranking is possible, the relative efficiency of quantiles estimation using ranked samples relative to SRS is high. This may translates to large savings in cost and time. Also, this conclusion holds even if the ranking is not perfect. Computer simulation results are given and real data from lows 65+ study is used to illustrate the method.

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An Efficient Representation of Edge Shapes in Topological Maps

  • Doh, Nakju Lett;Chung, Wan-Kyun
    • ETRI Journal
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    • v.29 no.5
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    • pp.655-666
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    • 2007
  • There are nodes and edges in a topological map. Node data has been used as a main source of information for the localization of mobile robots. In contrast, edge data is regarded as a minor source of information, and it has been used in an intuitive and heuristic way. However, edge data also can be used as a good source of information and provide a way to use edge data efficiently. For that purpose, we define a data format which describes the shape of an edge. This format is called local generalized Voronoi graph's angle (LGA). However, the LGA is constituted of too many samples; therefore, real time localization cannot be performed. To reduce the number of samples, we propose a compression method which utilizes wavelet transformation. This method abstracts the LGA by key factors using far fewer samples than the LGA. Experiments show that the LGA accurately describes the shape of the edges and that the key factors preserve most information of the LGA while reducing the number of samples.

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Organic Precipitate Flotation of Trace Metallic Elements with Ammonium Pyrrolidinedithiocarbamate (II). Application of Solvent Sublation for Determination of Trace Cd, Co, Cu and Ni in Water Samples

  • 김영상;정용준;최희선
    • Bulletin of the Korean Chemical Society
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    • v.19 no.1
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    • pp.50-56
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    • 1998
  • A solvent sublation was studied for the determination of trace Cd, Co, Cu and Ni in water samples. Ammonium pyrrolidine dithiocarbamate (APDC) was used as a complexing agent. Experimental conditions such as pH of solution, amounts of APDC, the type and amount of surfactant, the type of solvent, etc. were optimized for the effective sublation of analytes. After metal-PDC complexes were formed in sample solutions of pH 2.5, the precipitate-type complexes were floated in a flotation cell with an aid of sodium lauryl sulfate as a surfactant and by bubbling with nitrogen gas. The precipitates were dissolved and separated into the surface layer of methyl iso-butyl ketone (MIBK). The analytes preconcentrated were determined by a graphite furnace atomic absorption spectrophotometry (GF-AAS). Extractability of each element was 88% for Cd(Ⅱ), 86% for Co(Ⅱ), 95% for Cu(Ⅱ) and 76% for Ni(Ⅱ), respectively. And this procedure was applied to the analysis of real samples. From the recoveries of more than 92%, it was concluded that this method could be simple and applicable for the determination of trace elements in various water samples of a large volume.

Automatic Generation of Training Character Samples for OCR Systems

  • Le, Ha;Kim, Soo-Hyung;Na, In-Seop;Do, Yen;Park, Sang-Cheol;Jeong, Sun-Hwa
    • International Journal of Contents
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    • v.8 no.3
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    • pp.83-93
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    • 2012
  • In this paper, we propose a novel method that automatically generates real character images to familiarize existing OCR systems with new fonts. At first, we generate synthetic character images using a simple degradation model. The synthetic data is used to train an OCR engine, and the trained OCR is used to recognize and label real character images that are segmented from ideal document images. Since the OCR engine is unable to recognize accurately all real character images, a substring matching method is employed to fix wrongly labeled characters by comparing two strings; one is the string grouped by recognized characters in an ideal document image, and the other is the ordered string of characters which we are considering to train and recognize. Based on our method, we build a system that automatically generates 2350 most common Korean and 117 alphanumeric characters from new fonts. The ideal document images used in the system are postal envelope images with characters printed in ascending order of their codes. The proposed system achieved a labeling accuracy of 99%. Therefore, we believe that our system is effective in facilitating the generation of numerous character samples to enhance the recognition rate of existing OCR systems for fonts that have never been trained.

Detection of Human Papillomavirus in Normal Oral Cavity in a Group of Pakistani Subjects using Real-Time PCR

  • Gichki, Abdul Samad;Buajeeb, Waranun;Doungudomdacha, Sombhun;Khovidhunkit, Siribang-On Pibooniyom
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2299-2304
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    • 2012
  • Since there is evidence that human papillomavirus (HPV) may play some role in oral carcinogenesis, we investigated the presence of HPV in a group of Pakistani subjects with normal oral cavity using real-time PCR analysis. Two-hundred patients attending the Dental Department, Sandaman Provincial Hospital, Balochistan, Pakistan, were recruited. After interview, oral epithelial cells were collected by scraping and subjected to DNA extraction. The HPV-positive DNA samples were further analyzed using primer sets specific for HPV-16 and -18. It was found that out of 200 DNA samples, 192 were PCR-positive for the ${\beta}$-globin gene and these were subsequently examined for the presence of HPV DNA. Among these, 47 (24.5%) were HPV-positive with the virus copy number ranged between 0.43-32 copies per 1 ${\mu}g$ of total DNA (9-99 copies per PCR reaction). There were 4 and 11 samples containing HPV-16 and -18, respectively. Additionally, one sample harbored both types of HPV. Among the investigated clinical parameters, smoking habit was associated with the presence of HPV (p = 0.001) while others indicated no significant association. The prevalence of HPV in normal oral cavity in our Pakistani subjects appears to be comparable to other studies. However, the association between the presence of HPV and smoking warrants further investigations whether both of these factors can cooperate in inducing oral cancer in this group of patients.

Validation of One-Step Real-Time RT-PCR Assay in Combination with Automated RNA Extraction for Rapid Detection and Quantitation of Hepatitis C Virus RNA for Routine Testing in Clinical Specimens

  • KIM BYOUNG-GUK;JEONG HYE-SUNG;BAEK SUN-YOUNG;SHIN JIN-HO;KIM JAE-OK;MIN KYUNG-IL;RYU SEUNG-REL;MIN BOK-SOON;KIM DO-KEUN;JEONG YONG-SEOK;PARK SUE-NIE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.595-602
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    • 2005
  • A one-step real-time quantitative RT-PCR assay in combination with automated RNA extraction was evaluated for routine testing of HCV RNA in the laboratory. Specific primers and probes were developed to detect 302 bp on 5'-UTR of HCV RNA. The assay was able to quantitate a dynamic linear range of $10^7-10^1$ HCV RNA copies/reaction ($R^2=0.997$). The synthetic HCV RNA standard of $1.84{\pm}0.1\;(mean{\pm}SD)$ copies developed in this study corresponded to 1 international unit (IU) of WHO International Standard for HCV RNA (96/790 I). The detection limit of the assay was 3 RNA copies/reaction (81 IU/ml) in plasma samples. The assay was comparable to the Amplicor HCV Monitor (Monitor) assay with correlation coefficient r=0.985, but was more sensitive than the Monitor assay. The assay could be completed within 3 h from RNA extraction to detection and data analysis for up to 32 samples. It allowed rapid RNA extraction, detection, and quantitation of HCV RNA in plasma samples. The method provided sufficient sensitivity and reproducibility and proved to be fast and labor-saving, so that it was suitable for high throughput HCV RNA test.