• Title/Summary/Keyword: ELISA antibody

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An Enzyme-Linked Immunosorbent Assay for Detection of Cooked Goat Meat (가열 염소육의 판별을 위한 효소면역측정법)

  • Kim, Hyun-Jung;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.538-543
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    • 2000
  • This study was conducted to develop an enzyme-linked immunosorbent assay(ELISA) for the determination of cooked goat meat. Muscle proteins were extracted from goat meat by heating at $98^{\circ}C$ for 15 min. Major thermostable(TS) protein, whose size and pI are 36 and 38 kDa and 4.5 respectively, were purified by DEAE-Sephadex A-50 and Sephadex G-75 column chromatography. The TS protein was immunized into rabbits in order to produce goat specific antibodies. Competitive indirect ELISA(ciELISA) was established by using the anti-TS antibody. The antibody showed high reactivity toward the TS antigen and the boiled goat meat extract but it did not show any reactivities toward extracts of boiled chicken, pork, lamb, and beef. Thus, this ciELISA developed in this study could be applicable to identify goat species from cooked meat.

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Antibody changes in paragonimiasis patients after praziquantel treatment as observed by ELISA and immunoblot (폐흡충증에서 치료 후 혈청내 특이 항체의 변화)

  • 조승열;김석일
    • Parasites, Hosts and Diseases
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    • v.27 no.1
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    • pp.15-22
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    • 1989
  • To observe antibody changes after praziquantel treatment in paragonimiasis, a total of 46 serum samples from 13 serologically diagnosed patients was collected for 4~28 months. The specific antibody (IgG) levels were measured by enzyme- linked immunosorbent assay (ELISA). All but one patient who needed retreatment became symptom-free within a week. Antibody levels were dropped near to or below a cut-off absorbance (abs.) of 0.25 in varying intervals from 4 to 18 months. Of 9 patients who were retested within 3 months, 5 revealed temporary elevation of antibody level. After the elevation, the levels be낙an to decline slowly to negative ranges. If treated earlier after symptoms developed, the temporary elevation did not occur and intervals to negative conversion were shorter. By sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) /immunoblot, antigen-antibody reactions in individual patient faded gradually without significant changes in reacting antigen bands.

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Immunological Characterization of Vibrio vulnificus isolated from Marine Environment (해양에서 분리한 Vibrio vulnificus의 면역학적 특성)

  • Jung, Cho-Rok;Jeon, You-Jin;Heo, Moon-Soo
    • Korean Journal of Environmental Biology
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    • v.19 no.4
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    • pp.302-312
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    • 2001
  • Immunoglobulin G was purified by 40% $(NH_4)_2SO_4$ precipitation, DEAE-Sephadex, Sephadex G-150 column chromatographies from rabbit antiserum against V. vulnificus ATCC 27562 O antigen and used for immunological test for V. vulnificus isolates. The profiles of cell lysate total protein and outer membrane protein from the isolates were analyzed by SDS-PAGE and densitometry. The overall profiles in all isolates were similar. Distict protein band was observed in comparison with V. parahaemolyticus. Western Blotting with rabbit Immunoglobulin G against cell lysates and OMP of V. vulnificus isolates showed a strong antigenic response to antigen 66, 60, 54, 48, 33 and 26 kDa which were common to all strains examined. The 26 kDa antigen showed V. vulnificus specific antigen in comparison with Vibrio parahaemolyticus. A sandwich enzyme-linked immunosorbent assay was developed by using rat anti-V. vulnificus ATCC 27562 polyclonal antibodies as capture antibody, a purified rabbit IgG antibody as detector antibody, and goat anti-rabbit IgG-alkaline phosphatase conjugate as developer antibody. When four V. vulnificus isolates were tested, the reactivity showed from 50 to 70% by sandwich ELISA.

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Serum Antibody Ig G and Ig M Titers for Opisthorchis felineus Correlate with Eggs in Faeces - a Comprehensive Study in Chuvash Republic, Russia

  • Emelianov, Vladimir U;Skvortsova, Tatiana;Mikhailova, Lilia V;Shamitova, Elena N
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.281-283
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    • 2016
  • The Cholangiocarcinoma is a. The risk of development of cholangiocarcinoma, generally a rare type of a liver tumor, increases during infection of Opisthorchiasis. For this reason the timely detection of Opisthorchiasis is important for Cholangiocarcinoma prevention. There are many studies which concern the detection of pathogenesis of Opisthorchis viverrini infection but a little known about Opisthorchis felineus. In this study we investigate a correlation of the eggs which are found in a faeces and are comparable with a serum Ig G and Ig M antibody level that were detected with ELISA test in a large group of patients. The result is showing positive correlation between evidence of the Opisthorchis felineus eggs that were found in a faeces and antibody Ig G and Ig M level in a serum. Moreover the combination of two methods can improve the Opisthorchiasis diagnostic: the serum antibody and faeces investigation of eggs.

Identification of the IL-1$\beta$ inhibitor in the febrile patient urine by anti-IL-1$\beta$ monoclonal antibody (Anti-IL-1$\beta$ 단일클론 항체를 이용해서 발열환자의 뇨중 IL-1$\beta$ inhibitor의 확인)

  • 남경수;배윤수;남명수;오은숙;박순희;최인성;정태화
    • YAKHAK HOEJI
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    • v.37 no.4
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    • pp.420-426
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    • 1993
  • To effectively purify of IL-1 inhibitor from human febrile urine, we have established monoclonal antibody that reacts with human recombinant interleukin l$\beta$(IL-1$\beta$). The antibody, designated ON-1, was highly specific to IL-1$\beta$ and no cross-reaction with other cytokines(IL-l$\alpha$ and IL-4) was observed. As the results of ELISA inhibition assay and Western blotting method, it was further identified that ON-1 had high binding specificity with IL-1$\beta$. IL-1 receptor binding material from febrile patient urine was effectively purified with affinity column chromatography which conjugated with ON-1. This urinary material inhibited the thymocyte proliferation in a dosedependent manner. IL-l$\beta$ induced thymocyte proliferation activity was inhibited to 67.3% at 6 $\mu\textrm{g}$ of the purified urinary material. The result may suggest that this urinary material the purified urinary material. The result may suggest that this urinary material will have antagonic effect on IL-1 action mechanism and act IL-l$\beta$ inhibitor.

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A Study on the Activity of Antibodies Immunreaction to the Antigen of Paragonimus westermani (폐흡충에 대한 면역항체의 면역반응 활성에 관한 연구)

  • Kim, Soo-Jin;Lee, Ji-Hyon
    • Applied Microscopy
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    • v.28 no.4
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    • pp.527-538
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    • 1998
  • This study was performed to investigate the activity of antibodies in the tissues of Paragonimus westermani at the different developmental stages. Enzyme-linked immunosorbent assay (ELISA) and Immunelectron microscopy (IEM) were applied, using the dog sera infected with metacercariae isolated from Cambaroides similis. These dog sera were obtained from 3rd to 96th week after infection by bleeding. The supernatants of homogenated worms for worm antigen were used. The worm tissues were embedded in Lowicryl HM 20 medium, treated with infected serum and protein A gold complex (particle size; 12 nm) and observed by electron microscope. In the pattern of antibody levels by ELISA test in all developemental worm antigens, the activity of antibody was very weak in the 3rd week, but strong in the worm antibody from 4th to 20th week after infection. Its activity was maintained even till 96th week. The antibody level of the L2th week worm antigen was higher than those of the 20th and 48th week worm antigens. Generally, many gold particles were observed on the secretory granules and the epithelial lamellae. Thus, it was concluded that the antigenic materials in the developmental worm tissues were especially concentrated on the secretory granules in the parenchymal tissues and the epithelial lamellae in the lumen of the caecum.

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Analysis on antibody titer of structural protein after vaccination against foot-and-mouth disease virus (Type O) in zoo animals (동물원 동물에서 백신접종 후 구제역바이러스(O형) 구조단백질 항체가 분석)

  • Lee, Hyun-Ho;Eo, Kyung-Yeon
    • Korean Journal of Veterinary Service
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    • v.45 no.2
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    • pp.125-131
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    • 2022
  • The purpose of this study was to examine antibody titers to structural protein (SP) of the foot-and-mouth disease (FMD) virus after vaccination in animals of the Seoul zoo. After the initial inoculation of FMD vaccine to the susceptible animals of the zoo, a total of 235 blood samples were collected from 42 species of zoo animals during treatment or necropsy. All samples were tested by using enzyme-linked immunosorbent assay (ELISA). The overall positive rate of SP antibodies against FMD virus was 94.0% (221/235). However, the positive rates varied according to animal species. The results of positive rates in 30 species were 100% but in 12 species were 50-94.7%. We showed that most animals that have received FMD vaccine in Seoul zoo have been reached to the level of herd immunity against FMD virus after the vaccination. To the best of our knowledge, this study would be the first report for monitoring the vaccine-induced SP antibody titers against FMD virus after vaccination in various zoo animal species in Korea.

Enzyme-Linked Immunosorbent Assays of Pseudomonas tolaasii, a Bacterial Brown Blotch Pathogen of Oyster Mushroom. (느타리버섯 세균성갈반병균 Pseudomonas tolaasii의 효소면역검출법)

  • 이향범;전낙범;손동화;유승헌
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.238-243
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    • 1998
  • For simple and rapid detection of Pseudomonas tolaasii (PT), a bacterial brown blotch pathogen of oyster mushroom, enzyme-linked immunosorbent assays (ELISA) were developed. To produce specific antibody, PT ($5{\times}10^7$ cfu) and Freund's adjuvant were subcutaneously immunized into rabbits several times. By using the antiserum showing the highest titer, we established noncompetitive and competitive ELISA's. Standard curves of the ELISA's showed that the detection limits were $2{\times}10^2$cfu/ml and $3{\times}10^2$cfu/ml, respectively When investigated by noncompetitive ELISA, cross reactivities of the anti-PT antibodies against P. agarici, P. reactans, and other fluorescent Pseudomonas spp. were very low (<1/10$^3$), but those against P. solanacearum, Erwinia chrysanthemi, Streptococcus mutans, Xanthomonas citri, and a fungus Fusarium oxysporum were almost none. However, when investigated by competitive ELISA, the reactivities against any other strains except PT were almost none. When the ELISA's were applied to 18 strains derived from mushrooms in order to identify PT, only 11 strains showing both pathogenicity and white line reactivity were obviously positive. These results showed that the ELISA's could be convenient tools to detect PT in accordance with existing methods.

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Production of the polyclonal subunit C protein antibody against Aggregatibacter actinomycetemcomitans cytolethal distending toxin

  • Lee, Su-Jeong;Park, So-Young;Ko, Sun-Young;Ryu, So-Hyun;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.38 no.sup2
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    • pp.335-342
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    • 2008
  • Purpose: Cytolethal distending toxin (CDT) considered as a key factor of localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis is composed of five open reading frames (ORFs). Among of them, the individual role of CdtA and CdtC is not clear; several reports presents that CDT is an AB2 toxin and they enters the host cell via clathrin-coated pits or through the interaction with GM3 ganglioside. So, CdtA, CdtC, or both seem to be required for the delivery of the CdtB protein into the host cell. Moreover, recombinant CDT was suggested as good vaccine material and antibody against CDT can be used for neutralization or for a detection kit. Materials and Methods: We constructed the pET28a-cdtC plasmid from Aggregatibacter actinomycetemcomitans Y4 by genomic DNA PCR and expressed in BL21 (DE3) Escherichia coli system. We obtained the antibody against the recombinant CdtC in mice system. Using the anti-CdtC antibody, we test the native CdtC detection by ELISA and Western Blotting and confirm the expression time of native CdtC protein during the growth phase of A. actinomycetemcomitans. Results: In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and generated the anti CdtC antibody against recombinant CdtC subunit expressed in E. coli system. Our anti CdtC antibody can be interacting with recombinant CdtC and native CDT in ELISA and Western system. Also, CDT holotoxin existed at 24h but not at 48h meaning that CDT holotoxin was assembled at specific time during the bacterial growth. Conclusion: In conclusion, we thought that our anti CdtC antibody could be used mucosal adjuvant or detection kit development, because it could interact with native CDT holotoxin.

Development of Monoclonal Antibodies for Diagnosis of Plasmodium vivax

  • Linh, Nguyen Thi Phuong;Park, Hyun;Lee, Jinyoung;Liu, Dong-Xu;Seo, Ga-Eun;Sohn, Hae-Jin;Han, Jin-Hee;Han, Eun-Taek;Shin, Ho-Joon;Yeo, Seon-Ju
    • Parasites, Hosts and Diseases
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    • v.55 no.6
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    • pp.623-630
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    • 2017
  • Plasmodium lactate dehydrogenase (pLDH) is a strong target antigen for the determination of infection with Plasmodium species specifically. However, a more effective antibody is needed because of the low sensitivity of the current antibody in many immunological diagnostic assays. In this study, recombinant Plasmodium vivax LDH (PvLDH) was experimentally constructed and expressed as a native antigen to develop an effective P. vivax-specific monoclonal antibody (mAb). Two mAbs (2CF5 and 1G10) were tested using ELISA and immunofluorescence assays (IFA), as both demonstrated reactivity against pLDH antigen. Of the 2 antibodies, 2CF5 was not able to detect P. falciparum, suggesting that it might possess P. vivax-specificity. The detection limit for a pair of 2 mAbs-linked sandwich ELISA was 31.3 ng/ml of the recombinant antigen. The P. vivax-specific performance of mAbs-linked ELISA was confirmed by in vitro-cultured P. falciparum and P. vivax-infected patient blood samples. In conclusion, the 2 new antibodies possessed the potential to detect P. vivax and will be useful in immunoassay.