• Title/Summary/Keyword: Immuno-strip

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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.

Enzyme-linked Immunosorbent Assay Strip Sensor for Rapid Detection of Staphylococcus aureus (Staphylococcus aureus 신속 검출을 위한 효소면역측정 스트립 센서)

  • Park, So Jung;Kim, Young-Kee
    • Applied Chemistry for Engineering
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    • v.22 no.5
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    • pp.522-525
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    • 2011
  • In this study, an established enzyme-linked immunosorbent assay and immuno-chromatography technique are combined to fabricate an immuno-strip sensor for the detection of S. aureus. The immuno-strip is manufactured by using four different functional membranes. The capture antibody is immobilized on the nitrocellulose membrane due to the high affinity and the capillary action through porous membranes induces a flow of sample. A colorimetric signal is appeared according to the enzyme reaction and is analyzed by the digital camera (qualitative analysis) and home-made image analysis software (quantitative analysis). Under the optimal conditions, samples with S. aureus in the range of $2.7{\times}10^4{\sim}2.7{\times}10^7CFU/mL$ can be detected by the colorimetric method within 30 min.

Optimization of Fabrication Conditions for Immunosensor Strip to Detect Escherichia coli O157 : H7 (Escherichia coli O157 : H7 탐지용 면역센서스트립 제작 조건 최적화)

  • Park, So-Jung;Kim, Young-Kee
    • KSBB Journal
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    • v.24 no.3
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    • pp.253-258
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    • 2009
  • In this study, the optimization of fabrication conditions was accomplished to make immuno-strip biosensor by the combination of enzyme linked immunosorbent assay (ELISA) and immuno-chromatographic strip techniques for the detection of Escherichia coli O157 : H7. Optimal fabrication conditions of capture antibody concentration, detection antibody concentration, and additive composition of running buffer solution were determined. Optimal concentration was determined as 1.0 mg/mL for both of capture antibody and detection antibody. A composition of 0.5% Tween20 and 3% BSA were selected as optimal additive for buffer solution to prevent non-specific binding.

First report of Cucumber mosaic virus in African Impatiens (Impatiens walleriana) in Korea

  • Choi, Seung Kook;Choi, Gug-Seoun;Kwon, Sun-Jung;Cho, In-Sook;Yoon, Ju-Yeon
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.341-345
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    • 2015
  • Virus-like symptoms including stunt, severe mosaic with malformation of leaves, fern-like leaves and abnormal petals were observed from an African impatiens (Impatiens walleriana) grown in a plant nursery in Icheon, Korea. Serological analysis using immuno-strip kits for viruses reported in African impatiens indicated that Cucumber mosaic virus (named CMV-Im) was a causal agent for the symptomatic African impatiens. Biological properties of CMV-Im were analyzed using responses of host plant species, suggesting that CMV-Im is a typical strain that belongs to CMV subgroup I. RT-PCR analysis verified CMV-Im infection from naturally infected African impatiens or mechanically inoculated some host species. Analysis of multiple alignments of CMV capsid protein (CP) sequences showed that CMV-Im shared high CP amino acids identities with other CMV strains. Phylogenetic tree analysis for the CP sequences of CMV-Im and representative CMV strains confirmed that CMV is a typical member of CMV subgroup I. To our knowledge, it is the first report of CMV in African impatiens in Korea.

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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Trends in Rapid Detection Methods for Food-borne pathogenic Microorganisms by Using New Technologies (신기술 이용 식중독균 신속검출법 개발 동향 분석)

  • Kim, Hyun-Joo;Kim, Yong-Soo;Chung, Myung-Sub;Oh, Deog-Hwan;Chun, Hyang-Sook;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.376-387
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    • 2010
  • Recently, speedy, convenient and easy detection technologies have been developed rapidly and on the contrary, studies on development of traditional detectors applying biochemical characteristics has gradually been decreased. This review examined trend in current studies on detection of food-borne pathogenic microorganisms in the fields of selective media, immuno-assay, Polymerase Chain Reaction (PCR), microarray, terahertz spectroscopy & imagination and so on. Most traditional methods to detect the organisms from food matrix rely on selective media and such a method have disadvantages like long time requirement and distinguishing one species only from each selective medium although they are highly economical. Various new convenient methods such as Enzyme Linked Immuno-sorbent Assay (ELISA), paper-strip kit, fluoroimmunoassay etc. have been developed. The most ideal method for detecting food-borne pathogenic microorganisms in foods should be accurate, convenient, rapid and economical. Additionally, it is needed that capabilities of quantitative analysis and automation to be applied to industries.

Clinical usefulness of rapid antigen test to detect respiratory syncytial virus infection (Respiratory syncytial virus 감염진단을 위한 신속항원검사의 유용성)

  • Kim, Hyung Su;Kim, Hee La;Park, Ki Hyung;Cho, Kyung Soon
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1071-1076
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    • 2008
  • Purpose : Respiratory syncytial virus (RSV) is the most frequent cause of lower respiratory infections in infants and young children. Early detection allows quarantining of infected inpatients to prevent nosocomial transmission and to choose a treatment. To achieve rapid reporting, to facilitate prompt antiviral therapy, and to avoid unnecessary use of antibiotics, an easy, rapid diagnostic method for RSV is needed. We evaluated a lateral flow immunochromatography (RSV Respi-Strip test) and EIA (Enzyme immuno assay) compared to RT-PCR. Methods : From April 2007 to March 2008, 112 consecutive respiratory specimens (nasopharyngeal aspirates, throat swabs, tracheal aspirates, sputum) from patients who were suffering from the clinical signs and symptoms of respiratory tract infection were enrolled in Busan. A total of 112 patients were tested with RSV Respi-Strip (Corio-BioConcept, Belgium), EIA, and RT-PCR at the same time. Results : Of the 112 specimens tested, the number of children who showed positive results at RT-PCR and Respi-Strip were 45 and 42, respectively. The Respi-Strip rapid antigen test had a sensitivity of 88% and a specificity of 94%. The positive and negative predictive values were 90% and 92%, respectively. The agreement was 83%. Conclusion : In our study, the rapid antigen test had as much sensitivity as any method for detection of RSV. The test has many advantages such as easy performance, simple interpretation, and rapid results. If the rapid antigen test is widely applied in the clinical setting, the may be useful for diagnostic and epidemiological studies of RSV infection.

Effect of Microwave Plasma on Sterilization of Acidovorax citrulli Infected Watermelon Seeds (저온플라즈마(Microwave Plasma)를 활용한 Acidovorax citrulli 감염 수박종자의 살균 효과 검정)

  • Kim, Sang Woo;Ju, Han Jun;Gwon, Byeong Heon;Adhikari, Mahesh;Kim, Hyun Seung;Park, Mi-Ri;Lee, Youn Su
    • Research in Plant Disease
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    • v.27 no.1
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    • pp.8-16
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    • 2021
  • This study was conducted to check the sterilization efficacy of microwave plasma (MWP) against the watermelon seeds infected with Acidovorax citrulli 11-251. Watermelon seeds were artificially vacuum inoculated to produce A. citrulli 11-251 infected seeds. Aac ImmunoStrip and scanning electron microscope (SEM) results suggests that, seeds (coat and endosperm) were infected under the concentration of 1×107/30 min. MWP sterlization process was carried out at 50 W (3 min, 5 min, and 10 min), 80 W (3 min, 5 min, and 10 min), and 100 W (3 min, 5 min, and 10 min). According to the results, MWP sterilized the artificially inoculated seed coats by 95.96% at 80 W/10 min and seed endosperms by 100% at 100 W/10 min respectively. Although, seeds were sterlized by MWP, germination rate of seeds were low as compared to non treated (negative control) seeds. Moreover, cell membrane of A. citrulli 11-251 was damaged while observed in SEM after sterilized with MWP. Further studies regarding the appropriate sterilization condition by MWP against A. citrulli infected seeds for germination will be conducted in our next study.

Research Trend of Cronobacter Species Detection Methods: A Review (Cronobacter Species의 검출에 관한 연구동향: 총설)

  • Kwon, Heejun;Kim, Myunghee
    • The Korean Journal of Food And Nutrition
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    • v.28 no.4
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    • pp.728-736
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    • 2015
  • Cronobacter species (Cronobacter spp.), previously known as Enterobacter sakazakii, are gram negative food borne pathogenic bacteria. They pose a very high risk of infection to neonates and immuno-compromised individuals and can affect the human central nervous system. Consequently, survivors often suffer from severe neurological impairment including hydrocephalus, quadriplegia, and developmental delays. Cronobacter spp. were not only isolated from plant food and products such as cereals, fruits, vegetables, legume products, herbs, and spices but also from animal source foods such as milk, meat, fish, and products made from these foods. Therefore, rapid detection of Cronobacter spp. is essential for food safety. Many detection methods have been developed since the Cronobacter spp. were first reported. However, the development of more rapid, sensitive, and easy-to-use detection methods for the Cronobacter spp. is required. In this review, our aim was to study and compare the available detection methods for Cronobacter spp., including culture-based, molecular biology-based, and immunology-based methods. This study will contribute to the development of new and rapid detection method for Cronobacter spp.

Risk Assessment and Evaluation of Bt-transgenic Rice : Responses of Misgurnus anguillicaudatus and Cyprinus carpio fed on Bt-transgenic Rice Variety (해충저항성 Bt벼의 환경위해성 평가 : 해충저항성 Bt벼가 미꾸리(Misgurnus anguillicaudatus) 및 잉어(Cyprinus carpio)에 미치는 영향)

  • Oh, Sung-Dug;Lee, Dae-Yong;Sohn, Soo-In;Lee, Ki-Jong;Ryu, Tae-Hun;Lee, Jang-Yong;Park, Beom-Seok;Kweon, Soon-Jong;Suh, Seok-Cheol;Park, Jong-Sug
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.570-577
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    • 2011
  • We developed insect-resistant GM rice(Bt transgenic rice) by inserting the mCry1Ac1 a modified gene from the soil bacterium, Bacillus thuringiensis. The Bt transgenic rice expressing the Bttoxin mCry1Ac1 was tested for the effects on survival of Misgurnus anguillicaudatus and Cyprinus carpio, commonly used as a model organism in ecotoxicological studies. M. anguillicaudatus and C. carpio fed 100% ground rice in suspension, using either Bt rice or non-GM counterpart rice(Nakdong). The Bt rice used for the test were confirmed to have the mCry1Ac1 gene expression by the immuno-strip and ELISA analysis. Feeding test showed that no significant differences in cumulative immobility and abnormal response of M. anguillicaudatus and C. carpio fed on between Bt rice and non-GM counterpart rice. The 96hr-LC50 values showed no difference between Bt rice(>1,000mg/L) and non-GM rice(>1,000mg/L). We concluded that there was no significant difference in toxicity for non-target organisms(M. anguillicaudatus and C. carpio) between Bt rice and non-GM counterparts.