• Title/Summary/Keyword: immuno-assay

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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.

Studies on the development of enzyme linked immuno-sorbent assay (ELISA) for hepatitis B surface antigen (HBsAg) by monoclonal antibodies of different affinity constants

  • Kim, Gye-Won;Hong, Sung-Youl;Shin, Soon-Cheon;Lee, Sung-Hee;Kim, Won-Bae
    • Archives of Pharmacal Research
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    • v.10 no.1
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    • pp.18-24
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    • 1987
  • Mouse monocolonal antibodies to Hepatitis B surface antien (HBsAg) were prepared and their functional capabilities tested by the method of solid phase enzyme linked immuno sorbent assay (ELISA). HBsAg binding studies inicated that one monoclonal antibody 6E-1-1 bound more HBsAg at a faster rate than the other monoclonal antibodies. Also, for the binding inhibition studies with the selected monoclonal antibody 6E-1-1, one monoclonal antibody 8D-3-6 didn't exhibit binding inhibition for HBsAg. Then, a simultaneous ELISA method was developed for the immunodiagnosis of HBsAg. Different combinations of two monoclonal antibodies as solid phase and horseradish peroxidase (HRPO) labeled phase were studied. The combination of monoclonal antibody of higher affinity constant (6E-1-1) immobilized in a solid phase and monoclonal antibody of lower affinity constant (8D-3-6) as a HRPO laeled phase was more sensitive when two monoclonal antibodies of different affinity constants for HBsAg were prepared.

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Dimethylnitrosamine-Induced Reduction in the Level of Poly-ADP-Ribosylation of Histone Proteins of Blood Lymphocytes - a Sensitive and Reliable Biomarker for Early Detection of Cancer

  • Kma, Lakhan;Sharan, Rajeshwar Nath
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.15
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    • pp.6429-6436
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    • 2014
  • Poly-ADP-ribosylation (PAR) is a post-translational modification of mainly chromosomal proteins. It is known to be strongly involved in several molecular events, including nucleosome-remodelling and carcinogenesis. In this investigation, it was attempted to evaluate PAR level as a reliable biomarker for early detection of cancer in blood lymphocyte histones. PAR of isolated histone proteins was monitored in normal and dimethylnitrosamine (DMN)-exposed mice tissues using a novel ELISA-based immuno-probe assay developed in our laboratory. An inverse relationship was found between the level of PAR and period of DMN exposure in various histone proteins of blood lymphocytes and spleen cells. With the increase in the DMN exposure period, there was reduction in the PAR level of individual histones in both cases. It was also observed that the decrease in the level of PAR of histones resulted in progressive relaxation of genomic DNA, perhaps triggering activation of genes that are involved in initiation of transformation. The observed effect of carcinogen on the PAR of blood lymphocyte histones provided us with a handy tool for monitoring biochemical or physiological status of individuals exposed to carcinogens without obtaining biopsies of cancerous tissues, which involves several medical and ethical issues. Obtaining blood from any patient and separating blood lymphocytes are routine medical practices involving virtually no medical intervention, post-procedure medical care or trauma to a patient. Moreover, the immuno-probe assay is very simple, sensitive, reliable and cost-effective. Therefore, combined with the ease of preparation of blood lymphocytes and the simplicity of the technique, immuno-probe assay of PAR has the potential to be applied for mass screening of cancer. It appears to be a promising step in the ultimate goal of making cancer detection simple, sensitive and reliable in the near future.

Studies on Immuno modulating Acitivity of Fermented Sophorae Radix Extract (고삼(苦參) 발효 추출물의 면역활성에 관한 연구)

  • Kim, Hyung-Seok;Han, Hyo-Sang;Lee, Young-Jong
    • The Korea Journal of Herbology
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    • v.26 no.2
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    • pp.17-23
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    • 2011
  • Objectives : This study aims at examining the effect of the fermentative extract of root of Sophorae Radix on the immuno-modulating activity. Methods : Cell viabilities were measured by MTT assay. Effect of SFS on nitric oxide(NO), hydrogen peroxide production from RAW 264.7 cells was accessed by Griess reagent assay. Effect of SFS on productions of inflammatory cytokines such as TNF-${\alpha}$, IL-6 in LPS-induced RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. Results : The results of the experiment are as follows. 1. As a result of carrying out MTT assay to check the cellular toxicity of the fermentative extract of Sophorae Radix. There was not any excessive toxicity to the macrophage when the fermentative extract of root of Sophorae Radix was treated in different concentrations. 2. The fermentative extract of Sophorae Radix increased the generation of hydrogen peroxide in the macrophage and significantly restored the suppression of the generation of the hydrogen peroxide in the macrophage induced by LPS. 3. The fermentative extract of Sophorae Radix reduced the generation of NO in the macrophage and significantly suppressed the increase of the generation of NO in the macrophage induced by LPS. 4. The fermentative extract of Sophorae Radix significantly decreased the amount of TNF-${\alpha}$ generated in the macrophage induced by LPS when it was $25{\mu}g/mL$ or higher. Conclusion : These results suggest that SFS has anti-inflammatory moiety related with its inhibition of NO, hydrogen peroxide, TNF-${\alpha}$, IL-6, in macrophage led by LPS.

The Comparisons between RIA and CIA in CA19-9 Monitoring for Diagnosis of Pancreaticobiliary Tumors

  • Jung, Ha-Seung;Kim, Dae-Sik
    • Biomedical Science Letters
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    • v.15 no.2
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    • pp.135-139
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    • 2009
  • The CA(carbohydrate antigen)19-9 is complex protein that can be used as an important marker which aids the clinical diagnosis and prognosis of various pancreaticobiliary tumors. However, it was also reported that there were some CA19-9 positive patients with benign disease as using RIA method. The purpose of this study is to evaluate the clinical usefulness of serum level of CA19-9 with RIA(radioimmuno assay), CIA(chemiluminescence immuno assay), and conventional liver function tests. The correlation between CIA and RIA in CA19-9 of pancreatobiliary disease was 0.9833(P<0.01). Also, the correlations between CIA and RIA in CA19-9 of benign and malignant pancreaticobiliary tumor patients was 0.8714(P<0.01) and 0.9727(P<0.01) respectively. The correlation between CA19-9 and ALP was 0.5140(P<0.01) and CEA was 0.3385(P<0.05) as using CIA. The measurement of serum CA19-9 levels by CIA method may be useful in differentiating patients with malignant disease from those with benign disease in pancreaticobiliary tumors.

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Monoclonal antibody-based enzyme immuno-slide assay (EISA) in the rapid diagnosis of Peste des petits ruminants of goats

  • Das, Kamol K.;Rahman, M.B.;Shil, N.K.;Rahman, Md Siddiqur;Jang, Hyung-Kwan;Song, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.33 no.1
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    • pp.1-6
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    • 2010
  • Monoclonal antibody (mAb)-based enzyme immune-slide assay (EISA) was used for the detection of Peste des petits ruminants (PPR) virus from field samples collected from a natural outbreak. The clinicopathological study was undertaken to diagnose the case primarily of PPR. Antigen was detected from discharges and faeces of infected goats and swabs of postmortem lesions prepared on glass slide or glass plate using acetone fixation. Nasal discharge collected at the early stage of disease course or lung is an appropriate ante- or postmortem sample for this technique, respectively. Convalescent polyclonal sera collected from recovered animals which were diagnosed as PPR by EISA showed high antibody titer against PPR by C-ELISA, demonstrating the satisfactory specificity of the test. Therefore, EISA is a sensitive and specific assay to confirm PPR infection both in field and laboratory conditions and especially suitable for developing country.

Analysis of the Chemical Constituents of Agaricus brasiliensis

  • Cho, Soo-Muk;Jang, Kab-Yeul;Park, Hong-Ju;Park, Jeong-Sik
    • Mycobiology
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    • v.36 no.1
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    • pp.50-54
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    • 2008
  • This study examined the chemical composition of A. blasiliensis and the chemical structural properties of an immuno-stimulating polysaccharide. The amino acids, free sugars, and organic acids by HPLC and fatty acids by GC were analyzed. The immuno-stimulating substance from A. blasiliensis was extracted with hot water and purified by ethanol precipitation. It underwent ion exchange chromatography on DEAE-cellulose and gel filtration on Toyopearl HW 65F. Through GP-HPLC, the substance was found to be homogeneous. Its chemical structure was determined by $^{13}C-NMR$. Fatty acids, organic acids, and sugar alcohol composition consisted exclusively of linoleic acid, fumaric acid and mannitol, respectively. The amino acids were mainly glutamic acid, glycine, and arginine. By $^{13}C-NMR$ analysis, the immuno-stimulating substance was identified as ${\beta}-(1{\rightarrow}3)\;(1{\rightarrow}6)$-glucan, composed of a backbone with $(1{\rightarrow}3)$-linked D-glucopyranosyl residues branching a $(1{\rightarrow}6)$-linked D-glucopyranosyl residue. The ${\beta}$-glucan from A. blasiliensis showed pronounced immuno-stimulating activity on the antibody-production ability of B-lymphocytes by the hemolytic suspension assay. In these results, A. blasiliensis was estimated to have potent pharmacological properties and potential nutritional values.

Development of One-Step Immuno-Chromatography Assay System for Salmonella typhimurium (Immuno-Chromatography 방법을 이용한 식중독균(Salmonella typhimurium) 1단계 분석시스템의 개발)

  • 백세환;이창우이창섭육순학
    • KSBB Journal
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    • v.11 no.4
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    • pp.420-430
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    • 1996
  • One-step immuno-chromatography assay system for heat-killed Salmonella typhimurium antigens was developed. Three major components used were a glass fiber membrane (placed at the bottom of the system) with an antibody (specific to the analyse, detection antibody)-gold conjugate deposited in a dry state on the surface, a nitrocellulose membrane (middle) with an antibody (also, specific to the analyse but recognized different epitome: capture antibody) and anti-detection antibody immobilized in spatially separated areas, and a cellulose membrane (top) as absorption pad. These membranes were partially superimposed such that a wicking of aqueous solution containing sample can continuously take place through membranes. Variables that affected the system performance were the concentration of capture antibody, the location on the membrane, inert protein used for blocking of the membrane and for carrying the sample, and the concentration of the gold conjugate. Under optimal conditions, within 15 minutes after absorption of a sample solution from the bottom of the system antigen-antibody complexes of sandwich type were formed on the membrane surface area with immobilized capture antibody and a color signal was generated in proportion to the analyse concentration. The minimum do tection limit of the analyse was $1{\times}106$ Salmonella cells/mL.

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