• Title/Summary/Keyword: Product Specificity

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Application of the rpoS Gene for Species-Specific Detection of Vibrio vulnificus by Real-Time PCR

  • Kim, Dong-Gyun;Ahn, Sun-Hee;Kim, Lyoung-Hwa;Park, Kee-Jai;Hong, Yong-Ki;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1841-1847
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    • 2008
  • Vibrio vulnificus is a causative agent of serious diseases in humans, resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for the V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1 g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium, using the rpoS gene in pure cultures and in infected oyster tissues.

Studies on diaminododecane Utilization by Bacteria Studies on Diaminododecane Utilization by Corynebacterium sp. DAD 2-3 (Diaminododecane 자화균에 관한 연구 제2보 Corynebacterium sp. DAD 2-3의 Diaminododecane자화에 관한 연구)

  • 이상준;이종근
    • Korean Journal of Microbiology
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    • v.21 no.4
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    • pp.191-196
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    • 1983
  • A Corynebacterium sp. capable of utilizing diaminododecane (DAD) were isolated from the soil by enrichment culture. Among 9 different kinds of substituted alkanes containing CN, $NH_2$, Cl, and SH groups (monoterminally or diterminally substituted) tested as carbon source, the isolate, designated as DAD 2-3, utilized DAD, putrescine dihydrochloride, dodecane and laurylamine. Dodecanethiol, thioanisole, decanedithiol, dicyanooctane, laurylcyanide,and dichlorodecane were not utilized. When emulgen 950 was added to the medium, the growth of DAD 2-3 was slightly accelerated. Isolate DAD 2-3 grown in the medium with DAD as carbon source formed .alpha.-ketoglutaric acid. Metabolic product of DAD 2-3 grown in a medium without nitrogen source was different from that of grown in a medium with $NH_4NO_3$. When glucose, putrescine, n-dodecane and other alkane derivatives were tested in place of DAD, isolate DAD 2-3 yielded products different from those they formed with DAD suggesting specificity of DAD as a carbon source.

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Framework for the Quantitative Evaluation of Media Arts (미디어아트의 정량적 평가체계에 대한 연구)

  • Chung, Shin-Young;Eune, Ju-Hyun
    • Archives of design research
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    • v.19 no.2 s.64
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    • pp.139-150
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    • 2006
  • The evaluation of art has been historically based on the thorough reliance on the subjectivity of beholders due to the fact that the production and appreciation of works of art is founded on the similar(subjective) value. There had been little attempt to reverse such tendency by creating an objective and quantitative checklist for evaluating a work of art. Centering on the evaluation of the Media Art, which increased dramatically in numbers since the late 1960s, this is an attempt in systematizing the quantitative evaluation of Media Art by reflecting the idea of subjective criticism as well as its medium specificity. As such, the criteria for the evaluation consist of media evaluation, work evaluation, appreciation evaluation, product evaluation and exhibition evaluation. The evaluators can be divided into the general public, the Media Art specialist and the curatorial staff, according to their experiences in dealing with the Media Art. Based on the result of the FGD, the weight has been added to the evaluation system according to each evaluation criteria to ensure the balance between the objective and subjective criticism.

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Mutation Analysis in β2-Adrenergic Receptor Gene by Single Strand Conformation Polymorphism (SSCP) and Denaturing High Performance Liquid Chromatography (DHPLC) (SSCP와 DHPLC에 의한 β2-교감신경수용체 유전자의 돌연변이 분석)

  • Park, Sang-Bum;Han, Sang-Man;Nam, Youn-Hyoung;Jang, Won-Cheoul
    • Analytical Science and Technology
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    • v.17 no.1
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    • pp.53-59
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    • 2004
  • Up to now, methods for the detection of genetic alterations as single strand conformation polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE) have been used. It is too labor-intensive and expensive to serve for routine analysis. Moreover, lower in its sensitivity and specificity being also strongly dependent on the experience of the investigater. To improve these problems, we analysed mutation of ${\beta}_2$-adrenergic receptor gene that controls bronchial asthma by denaturing high performance liquid chromatography (DHPLC) according to ion-pair reversed phase chromatography (IP-RPC). We extracted genomic DNA from 80 asthma patients and then amplified DNA using PCR and analysed PCR product by SSCP and DHPLC. As a result, we analysed mutation frequency is 19 (23.75%) on SSCP and 25 (31.25%) on DHPLC in ${\beta}_2$-adrenergic receptor gene. We conclude that DHPLC is a fast and simple screening method rather than SSCP analysis.

Qualitative PCR Detection of vitamin E-enriched GM Perilla (비타민 E 강화 유전자변형 들깨에 대한 정성 PCR 분석법)

  • Kim, Jae-Hwan;Ahn, Ji-Hye;Song, Hee-Sung;Kim, Kyung-Hwan;Kim, Dong-Hern;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.192-195
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    • 2006
  • For the development of a qualitative PCR detection method for genetically modified perilla (Perilla frutescens), perilla species-specific gene, KAS-I (Beta-ketoacyl-ACP synthase I), was selected and validated as suitable for the use as an endogenous reference gene in perilla. Primer specificity was first tested by the means of qualitative PCR analysis. The primer pair Pfru3-F/R amplifying the perilla endogenous gene, KAS-I, gave rise to an amplicon 95 bp. No amplified product was observed when DNA samples from 15 different plants were used as templates. Qualitative PCR detection method was assayed with vitamin E-enriched GM Perilla developed in Korea. For the qualitative PCR detection method, the construct-specific detection primer pairs were constructed. The primer pair TMTO-F/R amplifying the junction region of TMT (${\gamma}$-tocopherol methyltransferase) gene and OCS (Octopine synthase) terminator introduced in GM perilla gave rise to an amplicon 148 bp.

Development of Simultaneous Analytical Method for Thiodicarb and its Metabolite Methomyl in Livestock Products (축산물 중 Thiodicarb와 대사산물 Methomyl의 동시분석법개발)

  • Chang, Hee-Ra;You, Jung-Sun;Ban, Sun-Woo;Gwak, Hye-min
    • Korean Journal of Environmental Agriculture
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    • v.40 no.2
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    • pp.142-147
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    • 2021
  • BACKGROUND: Agricultural use and pest control purposes of pesticides may lead to livestock products contamination. Thiodicarb and its degraded product, methomyl, are carbamate insecticides that protect soya bean, maize, fruit, and vegetables and control flies in animal and poultry farms. For maximum residue limit enforcement and monitoring, the JMPR residue definition of thiodicarb in animal products is the sum of thiodicarb and methomyl, expressed as methomyl. This residue definition was set to consider the fact that thiodicarb was readily degraded to methomyl in animal commodities. And therefore the simultaneous analytical method of thiodicarb and methomyl is required for monitoring in livestock products. METHODS AND RESULTS: The study was conducted using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) method and HPLC-MS/MS to determine the thiodicarb and methomyl in livestock products. The limit of quantitation (LOQ) was 0.01 mg/kg for livestock products, including beef, pork, chicken, milk, and egg. The coefficient of determinations (r2) for the calibration curve were > 0.99, which was acceptable values for linearity. Average recoveries at spiked levels (LOQ, 10LOQ, and 50LOQ, n=5) in triplicate ranged from 73.2% to 102.1% and relative standard deviations (RSDs) were less than 10% in all matrices. CONCLUSION: The analytical method was validated for the performance parameters (specificity, linearity, accuracy, and precision) in livestock products to be acceptable by the CODEX guidelines.

Development and Validation of an LC-MS/MS Method for Determination of Damaurone D in Rat Plasma and its Application to Pharmacokinetic Study in Rats

  • Lee, Wonpyo;Song, Im-Sook;Han, Young Taek;Choi, Min-Koo
    • Mass Spectrometry Letters
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    • v.10 no.1
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    • pp.38-42
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    • 2019
  • Damaurone D belongs to the genus Rosa and is a traditional medicinal product used for the treatment of depression, inflammation, and infectious diseases. The purpose of this study was to develop a simple liquid chromatography-tandem mass spectrometry method for the detection of damaurone D in rat plasma and to demonstrate its application in pharmacokinetic studies. Damaurone D and berberine (internal standard) were extracted with acetonitrile using a protein precipitation method. Mass transition was monitored in multiple reaction monitoring mode at m/z $323.2{\rightarrow}267.0$ for damaurone D and m/z $336.1{\rightarrow}320.0$ for berberine in positive ion mode. Analytical validation was conducted by evaluating the specificity, linearity, accuracy, precision, matrix effect, extraction recovery, and stability. The calibration curves were linear over 2-1000 ng/mL. The intra- and inter-day precision and accuracy of quality control samples were 4.79-13.33% and 86.23-102.75%, respectively. The matrix effect and extraction recovery were 96.11-98.47% and 96.11-102.25%, respectively. In the pharmacokinetic study after intravenous administration of damaurone D at a dose of 3 mg/kg in rats, the area under the curve and clearance of damaurone D in rat plasma were $16750.26{\pm}2676.10min{\cdot}ng/mL$ and $182.44{\pm}31.36mL/min/kg$, respectively.

Development and validation of an analytical method for the quantification of 2,6-diisopropylnaphthalene in agricultural products using GC-MS/MS

  • Lee, Han Sol;Park, Ji-Su;Lee, Su Jung;Shin, Hye-Sun;Chung, Yun mi;Choi, Ha na;Yun, Sang Soon;Jung, Yong-hyun;Oh, Jae-Ho
    • Analytical Science and Technology
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    • v.34 no.1
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    • pp.1-8
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    • 2021
  • An analytical method was developed and optimized for the quantification of a plant growth regulator, 2,6-diisopropylnaphthalene (2,6-DIPN), in agricultural products using gas chromatography-tandem mass spectrometry. The samples were extracted, partitioned, and were purified using a Florisil® cartridge. To validate the analytical method, its specificity, linearity, limit of detection (LOD) and limit of quantification (LOQ) of the instrument, LOQ of the analytical method (MLOQ), accuracy, and repeatability were considered. The method displayed excellent results during validation, and is suitable for the determination and quantification of the low residual levels of the analyte in the agricultural samples. All of the results with the optimized method were satisfactory and within the criteria ranges requested in the Codex Alimentarius Commission guidelines and the Ministry of Food and Drug Safety guidelines for pesticide residue analysis. The developed method is simple and accurate and can be used as a basis for safety management of 2,6-DIPN.

Formation of the Strategy of Digital Marketing of the Enterprise in the Conditions of the Competitiveness Intensification in the International Market

  • Solntsev, Sergii;Smerichevskyi, Serhii;Skyba, Halyna;Zabashtanska, Tetiana;Bazaliyska, Natalia;Kolbushkin, Yuriy
    • International Journal of Computer Science & Network Security
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    • v.22 no.2
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    • pp.47-56
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    • 2022
  • The article defines the principles of formation of digital marketing strategy of enterprises in the conditions of intensification of competition on the international market. The stages of development of digital marketing strategy of enterprises in the conditions of intensification of competition in the international market are substantiated, which includes: setting goals, which envisages observance of the principles of SMART-scheme; product or service analysis; monitoring of competitors; analytics of definition and segmentation of the target audience of the enterprise; selection of digital marketing tools and channels for promotion on the international market of products or services; formation of a unique, unique trade offer, selection of indicators for evaluating the effectiveness of digital marketing strategy and its tools. It is proved that according to the principle of SMART method of goal setting it is necessary that the goals have: specificity, measurability, achievability, relevance, achievement of the goal should be limited in time, have specific deadlines. To increase the effectiveness of digital marketing strategy, it is necessary to analyze the internal and external environment using the method of SWOT-analysis, the advantage of which is a comprehensive assessment of the company, competitors and the industry as a whole in the face of competition in the international market. The main indicators of evaluation of the effectiveness of digital marketing strategy in the conditions of intensification of competition on the international market are substantiated.

Evaluation of Uricult Trio Test as a Rapid Screening of UTI in Children with Fever (소아 요로감염 진단에 있어 신속배양검사(Uricult Trio)의 유용성에 관한 연구)

  • Lee, Jin;Kim, Eun Jin;Lee, Taek Jin;Chang, Jin Keun;Cha, Sung-Ho
    • Pediatric Infection and Vaccine
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    • v.17 no.2
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    • pp.74-82
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    • 2010
  • Purpose : The Dipslide culture test is a rapid method for diagnosis of urinary tract infection (UTI). The aim of this study is to determine the diagnostic availability of a urine Dipslide test for evaluation of UTI in febrile children. Methods : Urine specimens from 151 febrile infants were inoculated by a routine blood agar urine culture method and the Dipslide test at the same time. Following incubation for 16-24 hours, the results of the Dipslide test were read at the next visit. Both results of Dipslide and those of routine culture were compared. Results : The mean age of subjects was 15${\pm}$10.6 months. There were 150 infants (99.3%) who had fever with a mean duration of 2.6${\pm}$2.6 days. Thirty two infants (21.2%) were diagnosed as having UTI. Sensitivity and specificity of Uricult Trio CLED medium were 59.4% and 84.8%, respectively. Sensitivity and specificity of Uricult Trio E. coli medium were 60.0% and 96.2%, respectively. The Pearson correlation coefficient between results of Uricult Trio CLED medium and urine culture was 0.438 (P=0.01). Correlation between results of Uricult Trio E. coli medium and urine culture was 0.617 (P=0.01). Conclusion : The Dipslide test requires only 16-24 hours with high specificity in terms of UTI caused by E. coli without the problems associated with specimen delay. This test seems to be helpful for exclusion of UTI in febrile infants and it may reduce unnecessary hospitalization and antibiotic use. However, further studies are required before the product can be recommended as a routine diagnostic tool.