• Title/Summary/Keyword: inhibition ELISA

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Inhibition Enzyme-Linked Immunosorbent Assay (ELSIA) for Rapid Dection of Pseudomonas fluorescens in Raw Milk using IgY (난황 laY 항체의 Inhibition Enzyme-Linked Immunosorbant Assay(ELISA)법을 이용한 원유내 Pseudomonas fluorscens의 신속 검출 방법 개발)

  • 이승배;최석호;백두연
    • Food Science of Animal Resources
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    • v.20 no.3
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    • pp.231-235
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    • 2000
  • 원유에 존재하는 Ps. fluorescens의 균수를 신속하게 측정하기 위한 Inhibition Enzyme Linked Immunosorbant Assay (ELSIA)를 개발하기 위해 Ps, fluorescens(KCTC 2344)을 산란계에 접종하여 Ps.fluorescens에 대한 anti-Ps. fluorescens IgY 항체를 생산하고 그 항체의 역기를 ELISA로 측정한 결과 32일까지 항체 역가가 증가하였으며, 분리된 IgY 항체의 titer는 1:128,000으로 나타났다. Anti-Ps.fluorescens IgY 항체에대한 교차반응을 조사한 결과 그람양성균 Lactococcus faecalis, Staphylococcus aureus 뿐만 아니라 그람음성 균인 Achrombacter sal-monisida, Escherichia coil와도 교차반응을 거의 하지 않는 것으로 나타났다. Anti-Ps. fluo-rescens IgY 항체를 가지고 Inhibition ELISA 방법으로 Ps. fluorescens 균수를 신속한 측정할 수 있는 표준곡선을 작성한 결과 Ps. fluorescens균을 5.0$\times$$10^4$cfu/ml부터 5.0$\times$$10^{8}$cfu/ml 측정할 수 있는 것으로 나타났다.

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Enzyme Immunoassay for the Sulfamethazine Residues in Pork Tissue

  • Park, Jun-Hong;Lim, Yoon-Kyu
    • Journal of Food Hygiene and Safety
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    • v.11 no.4
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    • pp.287-290
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    • 1996
  • To control the maximum residue level (MRL) for sulfamethazine (SMZ) residues in pork tissue, a microbial inhibition method is a regulatory screening assay method in Korea. Microwell plate-based competitive enzyme immunoassay (ELISA) kit is avalable for routine screening of SMZ residues in pork tissue. One ELISA kit is evaluated. Phosphate buffer extracts of samples fortified with SMZ at 0, 1, 5, and 10 ng/g were used in a recovery test of the kit. Market pork samples were assayed by the kit. Recovery of sulfamethazine was 104% at 10 ng/g. Intraassay variations and interassay variations for the kit were 7.70% and 5.76%, respectively. Concentration causing 50% inhibition of color development compared with blanks was 16.4ng. The violative pork samples with over MRL (0.1 $\mu\textrm{g}$/g) was 4 of 32 cases (12.5%) by used ELISA kit. This result indicates a possibility of the ELISA kit for screening test of SMZ residues in pork tissue, and still needs a comfirmatory assay for mandatory purposes.

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Serodiagnosis of human paragonimiasis by ELISA-inhibition test using monoclonal antibodies (단클론항체를 이용한 폐흡충증의 면역진단)

  • Yong, Tae-Sun;Seo, Jang-Hun;Yeo, In-Seok
    • Parasites, Hosts and Diseases
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    • v.31 no.2
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    • pp.141-148
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    • 1993
  • ELISA-inhibition test using Paragonimus westermani specific monoclonal antibody (Mab) was investigated to improve the diagnostic specificity of paragonimiasis. By cell fusion, one hybridoma clone secreting un-n westemanl specific Mab was selected (Pwa-14), which reacted on bands of 28 kDa, 42.5 kDa, 89 kDa and 120.5 kDa. IFA showed Pwa-14 was located at the vitelline follicles. By micro-ELISA, 100% of 22 paragonimiasis cases were found positive, but 5 of 40 clonorchlasls cases (12.5%),3 of 26 cystlcercosis cases (7.7%) showed false positive. None of 10 sparganosis patients or 28 normal controls reacted positively. On the other hand, by ELISA-Inhibition test using a R westermcni specific Mab, 100% of patagonimlasls cases were found positive, and there were no positive in cysticercosis, sparganosis cases or normal controls, except 2 (5.0%) false-positive sera of 40 clonorchiasis cases. The ELISA-Inhlbltlon test using a Mab showed higher specificity in comparison with macro-ELISA for serodlgnosis of human paragonimlasis.

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Vinblastine Determination Measured by a Sensitive ELISA Inhibition Assay (ELISA Inhibition Assay에 의한 Vinblastine의 측정)

  • Jae Wha kim;Mi Young Han;Hee Gu Lee;Eun Young Song;Tai Wha Chung;Kyung Soo Nam;In Seong Choe
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.121-126
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    • 1996
  • Specific monoclonal antibodies(mAbs) against bis-indole alkaloids related to vinblastine were established to develop a simple and specific immunoassay system for the quantitation of Vinca alkaloids. Vinca alkaloids were extracted from tissue cultured cells of periwinkle plant (Vinca rosea L.). Spleen cells from Balb/c mice immunized with vinblastine-bovine serum albumin(VBL-BSA) conjugate as immunogen were fused with myeloma cells(Sp2/0-Ag.14) in the presence of polyethylene glycol. In the preliminary experiments, 32 clones which highly reacted with VBL-BSA conjugate were selected by ELISA(Enzyme-linked immunosorbent assay).These clones were further analyzed by inhibition assay of ELISA. The results obtained with two typical monoclonal antibodies, KN-1 and KN-2, were described. KN-1 exhibited considerable reactivities with soluble dimeric bis-indole alkaloids, whereas no cross reacted with monomeric bis-indole alkaloids. However KN-2 showed cross reactivity with mono- and di-meric bis-indole alkaloids. Furthermore, KN-1 was applied to the immunoassay system for determining the VBL amounts of in vitro cultured cell extracts. This assay system could detect dimeric vinca alkaloid as low as 0.05 nM.

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Serodiagnosis of cysticercosis by ELISA-inhibition test using monoclonal antibodies (단세포군항체를 이용한 효소면역억제측정 법에 의한 유구낭미충증의 혈청학적 진단)

  • 용태순;여인석
    • Parasites, Hosts and Diseases
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    • v.31 no.2
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    • pp.149-156
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    • 1993
  • Monoclonal antibodies (Mabs) were produced against crude scolex extract of T solium metacestodes, and applied to ELISA-inhibition test for improving the specificity of serodiagnosis of human cysticercosis. Four hybridomas secreting species-specific anti- cysticercal Mabs (Cya-1, Cya-7, Cya-28 and Cya-31) were selected. Each Mab reacted on antigenic components of 25.5 kDa (Cya-1), 28 kDa (rcya-7), 87.5 kDa (Cya-281), and 12.5 kDa (Cya-31). IFA showed that Cya-1 was located at the calcium corpuscles, and Cya-7 at the loose connective tissue of T soLium metacestode scolex. Cya-28 and Cya-31 reacted on the tegument of the scolex. By conventional ELISA, 23 out of 28 (82.1%) cysticercosis patients were found serologically positive, but 1 out of 9 (11.1%) sparganosls cases and 6 out of 31 (19.4%) paragonlmiasis cases showed false positives. By ELISA-Inhibition test using species-specific anti-cysticercal Mab Cya-7, 19 out of 28 (67.9%) cystlcercosis cases were found serologically positive, but there were no false positives In other parasitic infections.

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Comparative study on avian influenza virus antibody titer by hemagglutination inhibition test and enzyme-linked immunosorbent assay in the mass zone layer (산란계 밀집지역에서 혈구응집억제반응과 효소면역측정법을 이용한 가금인플루엔자 혈중항체가의 비교 조사)

  • 이정원;엄성심;이성재;서이원;서석열;정동석;송희종
    • Korean Journal of Veterinary Service
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    • v.27 no.1
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    • pp.81-87
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    • 2004
  • This study was conducted to investigate the similarity between hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay(ELISA), the HI titer and mean ratio S/P ratio) of avian influenza virus. To perform this study, the 1,457 sera of layers 21 farms in May, July and September, respectively. As a result of HI test, positive rates were 480 to 422 (92.1%) in May, 494 to 394(79.8%) in July and 483 to 402(83.2%) in September, and the mean antibody titer were 4.6, 4.3, 4.0 to 0.3 decreased, respectively. The positive rates by ELISA, 480 to 475(99.0%) in May, 494 to 485(98.2%) in July, 483 to 472(97.7%) in September, and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. The result of HI test and ELISA positive 480 to 422(92.1%), 475(99.0%), 494 to 394(79.8%), 485(98.2%) and 483 to 402(83.2%), 472(97.7%). Therefore, ELISA was shown more sensitive compare the HI titers.

The Reactivity of Antiserum Raised against Native Glucose-6-phosphate Dehydrogenase with Denatured Glucose-6-phosphate Dehydrogenase in Competitive ELISA

  • Kim, Moon-Hee
    • BMB Reports
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    • v.31 no.5
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    • pp.519-523
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    • 1998
  • We have previously reported that anti-glucose-6-phosphate dehydrogenase (G6PD) serum raised against native G6PD (nG6PD) enzyme recognized nG6PD antigen poorly in competitive enzyme-linked immunosorbent assay (ELISA) (Kim, 1997). In the present study, we investigated whether anti-G6PD serum raised against nG6PD can react with denatured G6PD effectively in competitive ELISA. We used partially active G6PD (paG6PD) by repeated freeze-thawing or SDS-denatured G6PD (SDS-G6PD) as both immobilized and soluble antigens, and anti-G6PD serum raised against nG6PD for competitive ELISA. The polystyrene cuvettes coated with either paG6PD or SDS-G6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of paG6PD or SDS-G6PD as competitors, followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA with paG6PD or SDS-G6PD antigen exhibited the sigmoidal dose-response curve characteristic of competition immunoassays. Furthermore, Triton-denatured G6PD (Triton-G6PD) was used in competitive ELISA. The paG6PD, SDS-G6PD, or Triton-G6PD used as competitors increased the inhibition of antibody binding to immobilized either of nG6PD or denatured G6PD compared with nG6PD competitor. The inhibition by denatured G6PD competitors was more pronounced at high competitor concentrations than at low counterparts. We conclude that anti-G6PD serum raised against nG6PD can effectively react with denatured G6PD in competitive ELISA and that our anti-G6PD serum recognizes denatured enzymes better than active enzymes.

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Correlations in the results of virus neutralization test, hemagglutination inhibition test, and enzyme-linked immunosorbent assay to determine infectious bronchitis virus vaccine potency

  • Park, Mi-Ja;Joh, Seong-Joon;Choi, Kang-Seuk;Kim, Aeran;Seo, Min-Goo;Song, Jae-Young;Yun, Seon-Jong
    • Korean Journal of Veterinary Research
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    • v.56 no.3
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    • pp.189-192
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    • 2016
  • The virus neutralization (VN) test was used to determine potency of the infectious bronchitis (IB) vaccine. The results of VN, hemagglutination inhibition (HI), and enzyme-linked immunosorbent assay (ELISA) were compared with those of the IBV M41. The $r^2$ values between VN and HI titers and the ELISA antibody titer were 0.8782 and 0.0336, respectively, indicating a high correlation between VN and HI, but not VN and ELISA. The Cohen's kappa coefficient between the VN titer of 2 $log_{10}$ and HI titer of 5 $log_2$ was 0.909. Our results showed that VN could be replaced with HI for testing the potency of IBV M41.

Comparison of serological methods for detection of avian influenza virus antibodies (가금인플루엔자 바이러스 항체검출을 위한 혈청학적 진단법 비교)

  • Han, Myung-guk;Park, Kyoung-yoon;Kwon, Yong-kuk;Kim, Jae-hong
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.73-80
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    • 2002
  • An enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin of swine influenza virus (H1N1) as antigen was developed for detection of antibody to avian influenza virus (AIV). The sensitivity and specificity of a developed and commercial available ELISA kits were compared with those of agar gel precipitation (AGP) test and hemagglutination inhibition (HI) test using sera collected from chickens under condition of field exposure. The concentration of antigen, serum dilution and concentration of enzyme-conjugated secondary antibody in developed ELISA (S-ELISA) were 0.5ug/100ul, 1:200 and 0.03ug/100ul, respectively. The correlation coefficients between S-ELISA and commercial ELISA and HI titers were 0.419 and 0.533, respectively. A significant correlation (p < 0.01) was not found between HI and ELISA titers. The S-ELISA was found to be as more sensitive and specific than the AGP test, showing 86.8% sensitivity and 85.3% specificity. It is suggested that the ELISA using the SIV as antigen may be useful method as an investigating tool for AIV serological surveillance.

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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