• Title/Summary/Keyword: Nitrocellulose

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An Enzyme-linked Immunosorbent Assay Strip Sensor for the Detection of Legionella Pneumophila (Legionella Pneumophila 검출을 위한 효소면역측정 스트립 센서)

  • Kim, Young-Kee;Park, Sojung
    • Applied Chemistry for Engineering
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    • v.25 no.5
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    • pp.544-547
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    • 2014
  • In this study, an enzyme-linked immunosorbent assay (ELISA) and immuno-chromatographic technique were combined to fabricate immuno-strip sensors for the detection of Legionella pneumophila. The immuno-strip sensor was manufactured with four different membranes. A nitrocellulose membrane was used to immobilize capture antibody and generate signals due to the high affinity to antibodies, and glass fiber membranes were used as a conjugate release pad and a sample application pad. A cellulose membrane was used as an absorption pad to induce sample flow by the capillarity. Colorimetric signals produced by sandwich immuno-reaction and enzyme reaction could be analyzed qualitatively and quantitatively within 30 min. Under the given experimental conditions, sensor signals with L. pneumophila samples were observed qualitatively by naked eyes and measured quantitatively in a range of $1.3{\times}10^3-1.3{\times}10^6CFU/mL$ with a digital camera and home-made image analysis software.

Development of Rapid Immune-gold Strip Kit for On-Site Diagnosis of Tomato spotted wilt virus

  • Yoon, Ju-Yeon;Choi, Gug-Seoun;Cho, In-Sook;Choi, Seung-Kook
    • Research in Plant Disease
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    • v.20 no.1
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    • pp.15-20
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    • 2014
  • A rapid, user-friendly and simple immune-chromatographic dipstick kit named 'rapid immune-gold strip' (RIGS) kit was developed in a novel single strip format to detect on-site detection of Tomato spotted wilt virus (TSWV). Immunoglobulin G (IgG) from polyclonal antisera raised in rabbits against TSWV was purified through protein-A affinity chromatography and then the purified TSWV-IgG was conjugated to colloidal gold nano-particles which served as a test line on nitrocellulose membrane. Protein A that non-specifically binds to TSWV antibody was used as a control line on the same strip. The diagnosis process with the TSWV-RIGS involves simply grinding the suspect plant sample in a bag that contains the extraction buffer and inserting the strip the bag. Results can be seen in 2-5 minutes. The flow of the complexes of gold particles coated with TSWV-IgG and a crude sap from TSWV-infected pepper, tobacco and tomato plants resulted in intensive color formed on the test lines proportional to the concentrations of TSWV. The RIGS-TSWV kit did not show any cross-reactions against other tomato-infecting viruses unrelated to TSWV. These results indicate that the TSWV-RIGS kit is highly sensitive and is not required for laboratory training and experience prior to testing. The TSWV-RIGS kit is suitable for on-site detection of suspect TSWV-infected plants as well as for laboratory diagnosis.

Task-Specific Hazardous Chemicals Used by Nail Shop Technicians (네일 샵 종사자들의 직무 형태별 취급 유해화학물질)

  • Choi, Sangjun;Park, Sung-Ae;Yoon, Chungsik;Kim, Sunju
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.25 no.4
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    • pp.446-464
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    • 2015
  • Objectives: This study was conducted to evaluate the task-specific hazards of chemicals used by nail technicians in Daegu Metropolitan City. Materials: A total of 30 nail shops located in Daegu City were surveyed to investigate the major tasks and practices performed by nail technicians and the ingredients listed in nail care products used in shops. We also collected instructions for use and material safety data sheets(MSDSs) of nail care products and compared CAS Nos. of ingredients with the lists of chemicals regulated by the Industrial Safety and Health Act(ISHA) and Chemical Substances Control Act(CSCA). Results: A total of 125 chemical ingredients were found in 468 nail care products used at the 30 nail shops. The most frequently found ingredients were ethyl acetate(72%), followed by n-butyl acetate(71.8%), isopropanol(56%), benzophenone(51.1%), nitrocellulose(46.4%) and ethanol(45.3%). Comparing six tasks, the task of manicuring used the most products at 222 products containing 91 ingredients. Among the 125 ingredients, there are 31 chemicals with occupational exposure limits(OEL) designated by the Ministry of Employment and Labor(MoEL), eight categorized as carcinogens, one mutagen and two reproductive toxic chemicals. In terms of carcinogens, formaldehyde was identified as the only confirmed human carcinogen(1A). We found that there was one chemical with a permissible limit, one special management substance, 18 workplace monitoring substances and ten special health diagnosis substances regulated by ISHA. For CSCA, nine poisonous substances, six substances requiring preparation for accidents and one restricted substance were identified. Conclusions: Based on these findings, formaldehyde was identified as one of the chemicals that should most strictly be controlled for the protection of the health of nail technicians and customers. At the same time, it is necessary to distribute materials with detailed hazardous information of nail care products for nail shop technicians.

Identification of Phosphoproteins Induced by AT1 Receptor Blocker Losartan (항고혈압 치료제 로사탄에 의해 인산화 되는 단백질 발굴)

  • Lee, Chang-Woo;Kim, Mi-Jin;Jang, Sei-Heon
    • Journal of Life Science
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    • v.18 no.7
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    • pp.918-923
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    • 2008
  • The angiotensin II receptor ($AT_1R$) antagonists are effective in treating patients with hypertension and showed beneficial effects in diabetes and other metabolic diseases. The beneficial effects of $AT_1R$ antagonists are mainly considered to be from inhibition of Ang $II-AT_1R$ signaling pathway such as the activation of NADPH oxidase and the generation of reactive oxygen species. In this study, we examined whether antagonist of the $AT_1R$ could account for phosphorylation of proteins in cells using antibody array. We have selected 6 proteins with Ser/Thr-phosphorylation sites and 12 proteins with Tyr-phosphorylation sites based on literature search. Upon $AT_1R$ antagonist losartan treatment to serum-starved COS-1 cells, there was ${\sim}20%$ increase of Ser phosphorylation in small GTPase RhoA. RhoA is known to be responsible for cytoskeleton rearrangement and is down-regulated upon Ser phosphorylation in vivo. Our finding provides a new insight into the mechanism and signaling pathway of the $AT_1R$ antagonist in cells.

Development of Membrane Strip Assay System for Lipoprotein Cholesterol Based on Liquid-Phase Enzyme Reactions (액상 효소반응을 이용한 Membrane Strip 형 Cholesterol 측정시스템의 개발)

  • 신인수;목락선;장미라;백세환
    • KSBB Journal
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    • v.13 no.5
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    • pp.577-584
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    • 1998
  • A sensitive membrane strip assay for plasma lipoprotein cholesterol that can be performed without handling reagents has been investigated. We previously developed an assay system with immobilized enzymes (cholesterol esterase and cholesterol oxidase) on the surfaces of nitrocellulose membrane(1). In such a case, the amount of enzymes present on the membrane was limited by its surface area and, thus, the detection capability was relatively poor (> 50 mg/dL cholesterol). To overcome this problem, we devised a new system with non-immobilized enzymes by placing them within interstitial spaces of a celullose membrane pad in a dry state. Upon contact with sample medium, the enzymes were immediately dissolved and participated in the reactions with cholesterol in a liquid phase. We constructed a user-friendly system consisting of four membrane pads fro sample application, cholesterol decomposition, color development as signal, and medium absorption to invoke a continuous flow (sequential location from the bottom). A sample containing lipoproteins was added into the application pad by capillary action and transferred to the next pad for decomposition. The decomposition pad (namely, enzyme pad) contained a detergent (sodium cholate) for the destruction of lipoprotein particles, the two enzymes for cholesterol decomposition, and a chromogen (3,3'-diaminobenzidine). As a consequence of the enzyme reactions, hydrogen peroxide was produced, and then reacted in the presence of the chromogen with horseradish peroxidase immobilized on the signal generation pad. Finally, a colorimetric signal directly proportional to the cholesterol concentration was produced. The detection limit determined from this system under optimal conditions was at least 2 times lower than of the enzyme-immobilized system.

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Stabilization of Dry Immuno-Diagnostic Kit with Immobilized Antibody and Determination of Its Shelf-Life (항체가 건조상태로 고정된 면역진단키트의 안정화와 Shelf-Life 결정)

  • 이창우;조정환;육순학;권오협;박영남;박세환
    • KSBB Journal
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    • v.13 no.5
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    • pp.502-510
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    • 1998
  • Stabilization of antibody, which is specific to Salmonella typhimurium antigens, present in dry states on membranes was accomplished, and its shelf-life, i.e., duration for maintaining minimum 90% of the initial activity, under optimal conditions was determined. To prepare two major components of an immuno-strip, the antibody was not only immobilized on nitrocellulose membrane surfaces but also placed within the pores of glass fiber membrane after conjugating it with old colloids as signal generator. Among potential stabilizers of the immuno-components, a disaccharide, trehalose, showed a significant protection effect of immunoglobulin structure from thermal energy. Optimal concentrations of trehalose for the respective component were significantly different (8-fold higher for the antibody-gold conjugate than for the immobilized antibody), which probably resulted from distinct densities and configurations of antibody present on the membranes. An additional requirement for the gold conjugate was freeze-drying of this substance such that the conjugate can be readily resolubilized upon contact with aqueous medium. By using the components prepared under optimal conditions, immuno-strips were constructed and exposed to thermal energy. Signals with less than 10% decrease in the intensity were maintained for approximately 21 days at 60$^{\circ}C$. Compared to previous reports, this result represented a 2-year shelf-life at room temperature. it was, however, two times longer if determined from thermal acceleration tests based on the theory of inactivation rate of protein. Such discrepancy between the two estimates could be mainly attributed to errors in accurately controlling temperatures and also to changes in the physical properties of membranes due to a high thermal energy.

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Detection of Waterborne Pathogens in Public Bath Houses by PCR-Reverse Blot Hybridization Assay (PCR-REBA) (분자생물학적 방법인 PCR-REBA를 이용한 대중목욕탕 수질 중 수인성병원성미생물 검출)

  • Song, Woon-Heung;Choi, Seung-Gu;Yang, Byoung-Seon;Lee, Jae-Sang
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.8
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    • pp.3517-3522
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    • 2011
  • Contamination of public bath water by waterborne pathogens can cause disease outbreaks and contribute to background rates of disease. The aim of this study is to determine the prevalence of waterborne pathogens in public baths. A total of 30 water samples were collected from 30 different public baths in seoul, Korea. Pathogens in water samples were concentrated by 0.45 ${\mu}m$ nitrocellulose membrane filter, analyzed by both cultivation and polymerase chain reaction-reverse blot hybridization (PCR-REBA) of partial 16S rRNA gene. Various microorganisms including Escherichia coli and Shigella spp. were identified by microbiological cultivation. E. coli, Shigella spp., Salmonella spp., Pseudomonas spp. and Mycobacterium spp. were identified by PCR-REBA. Our results suggest that appropriate hygiene practice and continuous monitoring is needed for reducing health risk associated with public bath houses.

Preparation and Coating of Red Colored Artificial Pearl by CVD Method (CVD법을 이용한 적색 인조진주 코팅 및 제조)

  • Shin, Cheol-Woo;Choi, Kyoung-Rim;Lee, Dong-Kyu
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.3
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    • pp.857-864
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    • 2018
  • Demand for developing artificial green pearl that meets the needs of modern people has been increasing. In this paper, eco-friendly inorganic pearlescent pigment was used instead of organic pigment and urethane resin was substituted for nitrocellulose which has been used as main materials in previous preparation method, increasing gloss from 73.4% to 86.7%. Urethane was coated on substrate before finishing with CVD, resulting high gloss of 96%. Colorimeteric analysis shows that a* and b* of CIE value was changed from +37.7 and +24.5 to +31.9 and +14.2 respectively because of CVD finishing, obtaining colorful, high gloss and durable artificial pearl. Quality and toxicity of samples was established by chemical resistance, glossiness, colorimeter, surface roughness, wear resistance, content of heavy-metal, and salt water test.

A Study on the Shelf-Life Prediction of the Domestic Single Base Propellants Ammunition : Based on 105mm High Explosive Propellants (국내 단기추진제 탄약의 저장수명 예측에 관한 연구 : 105미리 고폭탄 추진체를 중심으로)

  • Choi, Myoungjin;Park, Hyungju;Yang, Jaekyung;Baek, Janghyun
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.37 no.3
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    • pp.36-42
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    • 2014
  • Domestic 105mm HE (High Explosive) shell is composed of three parts that are Fuze, Projectile and Propellants. Among three parts, propelling charge of propellants part consists of single base propellants. It has been known that the lifespan of single base propellants is affected by a storage period. These are because Nitrocellulose (NC) which is the main component of propelling gunpowder can be naturally decomposed to unstable substances similar with other nitric acid ester. Even though it cannot be prevented fundamentally from being disassembled, a decomposition product ($NO_2$, $NO_3$, and $HNO_3$) and tranquillizer DPA (Diphenylamine) having high reactivity are added into a propellant to restrain induction of automatic catalysis by a decomposition product. The decay rate of the tranquillizer is also affected by a production rate of the decomposition product of NC. Therefore, an accurate prediction of the Self-Life is required to ensure against risks such as explosion. Hereupon, this paper presents a new methodology to estimate the shelf-life of single base propellants using data of ASRP (Ammunition Stockpile Reliability Program) to domestic 105mm HE (propelling charge of propellants part). We selected four attributes that are inferred to have influence on distribution of the DPA amount in a propellant from the ASRP dataset through data mining processes. Then the selected attributes were used as independent variables in a regression analysis in order to estimate the shelf-life of single base propellants.

Development of a Method for Rapid Analysis of DNA Hybridization (측방유동방식 신속 DNA 교잡 분석법의 개발)

  • 정동석;최의열
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.114-117
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    • 2003
  • In molecular biology, it is necessary to develop an easy and rapid method to identify a specific DNA sequence. Though Southern and Northern blot techniques have been used widely for the analysis of gene structure and function, those methods are inconvenient in the points that we need to control incubation temperature, time, and other parameters to get the final result. In this study, we report a new method for the rapid analysis of specific DNA sequence with the modification of an immunochromatographic method. The lateral flow DNA analysis strip is composed of a sample pad, a nitrocellulose membrane for the separation and propagation of analytes, and an absorption pad for the generation of capillary action. Capture DNA was immobilized on the membrane by UV cross-linking and target DNA was labeled with Cy-5 for signaling. The samples containing target DNA were applied onto the sample pad, incubated for 15 min for separation, and scanned with a GSI fluorescence scanner. Though the hybridization reaction occurs in a short time without any washing steps, there appears to be little cross hybridization between the different sequences. The result showed a possibility that the new method can be used for the rapid identification of specific DNA sequence among the samples.