• Title/Summary/Keyword: ELISA antibody

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Comparison between indirect immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to porcine reproductive and respiratory syndrome virus(PRRSV) (돼지 생식기호흡기증후군 바이러스항체 검색에 있어 간접형광항체법(IFA) 과 효소면역법(ELISA)의 진단효율 비교)

  • Park, Choi-kyu;Lyoo, Young-soo;Lee, Chang-hee;Jung, Jong-wook
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.314-318
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    • 1998
  • An establishment of effective control measures to PRRSV infection in swine industry depends on a sensitive and specific diagnosis to detect either viral antigen and/or antibodies to PRRSV. Several diagnostic methods are available to detect antibodies against PRRSV, including IPMA, IFA and ELISA tests have been successfully developed. Sensitivity of the indirect immunofluorescent assay in MA-104 cells using Korean field isolate PL96-1 was superior to that of VR-2332 and field isolate PL96-2. Sensitivity and specificity of the IFA test with PL96-1 were comparable to those of commercial ELISA test kit but ELISA test was more sensitive for the detection of declining antibodies to PRRSV in finishing pigs. In this study we concluded that IFA and ELISA test could be utilized to detect antibodies to PRRSV and the results generated from these two tests were comparable and there were no significant difference between these two tests.

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Establishment of Indirect Competitive ELISA for the Detection of Zearalenone Produced by Fusarium sp. (Fusarium속이 생성하는 zearalenone 측정을 위한 Indirect Competitive ELISA의 확립)

  • 강성조;정덕화
    • Journal of Food Hygiene and Safety
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    • v.13 no.4
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    • pp.419-424
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    • 1998
  • An enzyme-linked immunosorbent assay (ELISA) was established for the detection of zearalenone by using monoclonal antibodies produced by Z-M-26 hybridoma cells when injected into a mouse and zearalenone-oxime-OV A conjugate. Zearalenone-oxime-OV A conjugates were diluted with carbonyl buffer, coated to 96 well microtiter plates at $4^{\circ}C$ overnight and blocked with 1% BSA overnight. One thousand times diluted antibody solution together with standard zearalenone or sample was added to 96-well microtiter plates and stood overnight. A secondary antibody conjugated with HRP was added and an hour later, enzyme substrate (TMBZ) solution was added for color develpment. Mter 30 minutes, coloring reaction was terminated by adding 2 N $H_2S0_4$ and the O.D. was measured at 450 nm. Detection range of this method was about 0.1~100 ppb. The established indirect competitive ELISA method was suitable for a rapid and effective analysis of zearalenone in agricultural products.

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Nanobiocatalyst-Linked Immunosorbent Assay(NBC-LISA) (나노바이오촉매 기반 효소결합면역흡착검사)

  • Lee, Inseon;Hwang, Sang Youn;Kim, Jungbae
    • Korean Chemical Engineering Research
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    • v.49 no.4
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    • pp.387-392
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    • 2011
  • Enzymes are being used in various fields due to their unique property of substrate specificity. Enzyme-linked immunosorbent assay(ELISA) has enabled the detection of various antigens by reporting the binding event of antigen and antibody via enzyme-catalyzed reaction. However, the sensitivity improvement of conventional ELISA has been limited because only one enzyme molecule is conjugated to one molecule of antibody. To overcome this limitation and further improve the sensitivity of ELISA, there have been efforts to increase the number ratio of enzymes to antibody. Recently, the nanobiocatalytic approaches, with their successful enzyme stabilization, improved the performance stability as well as sensitivity in a modified protocol of ELISA. The present paper introduces the basic principle of ELISA, and the recent efforts to improve sensitivity and performance stability of ELISA by using the nanobiocatalytic approaches.

Serological Cross-Reactivity between Sarcocystis and Toxoplasma in Pigs (돼지에 있어서 Sareocustis와 Toxoplusma 감량의 혈청학적 교차반응 시험)

  • 문무홍
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.188-194
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    • 1987
  • The development of antibody titers and crossreaction between Sarcocystis and Toxoplasma were investigated by means of IF A test and ELISA in pigs experimentally infected with $1.5{\times}10^6$ S. suicanis sporocysts and 10,000 T. gondii oocysts, respectively. The intact and soluble Sarcocystis antigens were prepared from the bradyzoites harvested by peptic digestion of infected pork. The intact and soluble Toxoplasma antigens were prepared from the tachyzoites in mouse peritoneal cavity. IgG antibodies in pigs infected with Sarcocystis and Toxoplasma, respectively were detected first at 2 weeks post infection on both IF A test and ELISA. The antibody titer to Toxoplasma reached its maximum at 6 weeks post infection and decreased thereafter. The antibody titer to Sarcocystis reached its maximum terminally. The cross-reaction titer in pigs infected with Toxoplasma against Sarcocystis antigen was up to 1 : 16 in IFA test and up to 1 : 32 in ELISA. The titer in control group was below 1 : 4 in both reactions.

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Comparative evaluation of indirect immunofluorescent antibody test with enzyme-linked immunosorbent assay in serodiagnosis of human neurocysticercosls (뇌낭미충증의 혈청학적 진단에 있어서 간접 형광항체 반응 및 효소연결성 면역흡착 검사의 비교 평가)

  • Eom, Gi-Seon;Jo, Seung-Yeol;Im, Han-Jong
    • Parasites, Hosts and Diseases
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    • v.26 no.1
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    • pp.27-32
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    • 1988
  • The applicability of indirect immunoftuorescent antibody test (IFAT) was compared with enzyme-linked immunosorbent assay (ELISA) in sera from 163 cases of confirmed neurocysticercosis, 101 other neurologic and parasitic diseases and 100 normal controls. As antigen, frozen sections of a Taenia solium metacestode from a human brain was used in IFAT and cystic fluid was used in ELISA. For the detection of specific IgG antibody, IFAT was equally sensitive (89.6%) and specific (85.1%) as ELISA. The antibody titers by IFAT were correspondingly increased with mean absorbance of ELISA. The corresponding rate of positivity in the two techniques was 90.8%. Except for the difficulty in detecting antibodies in cerebrospinal fluid (CSF), IFAT was concluded to be very useful for the serodiagnosis of human neurocysticercosis.

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Polyclonal Antibody against Paenibacillus larvae and its Application (Paenibacillus larvae에 대한 다클론 항체 및 그 응용)

  • 백경찬;양옥순;정규회;윤병수
    • Korean journal of applied entomology
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    • v.41 no.1
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    • pp.49-53
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    • 2002
  • Paenibacillus larvae is a gram-positive, spore-forming bacterium that is etiological agent for american foulbrood disease (AFB), which is the most severe disease in honey bee. To detect P. larvae from infected honeybee-comb or larvae, polyclonal antibody against whole bacterium was produced from guineapig and its specificity was evaluated. After optimization of ELISA-based detection system using these antibodies, a number of different P. larvae strains were analysed. Polyclonal antibody against P. larvae ATCC 25747 showed high affinity to most strains of P. larvae including P. larvae. strain ATCC 9545 (type strain), ATCC 25747 and other korean strain, SJl5 but exhibited no cross-reaction with other bacterial species. Additionally, this type of ELISA system was used for the detection of AFB in field-application The results have shown that this antibody could be useful for the rapid identification and monitoring of P. larvae in honeybee-comb.

The serological studies on infectious bursal disease (전염성 F낭병에 대한 혈청학적 연구)

  • 정영미;서석열;도홍기;조정곤;노수일
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.271-279
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    • 2000
  • This study was carried out to provide the fundamental information for development of proper vaccination program against infectious bursal disease(IBD) to the local chicken farms. The antigen detection was peformed from 8 samples of bursa of Fabricius with agar gel precipitation(AGP) and indirect immunofluorescent assay(IFA), And also, the antibodies in serum samples were detected by the various serological methods such as commercial ELISA assay, AGP and virus neutralization(VN) test. 1. The antigen detection rates were 25% for AGP which is 2 out of 8 farms and 10 out of 40 bursas, and 25% which Is 2 out of 8 farms and 20% 8 out of 40 bursas for IFA, respectively. 2. The mean titer of maternal antibody (>3,000) existed until 10 days of the age with ELISA-GMT. 3. The antibody positive rates which are over 80% showed until 5 days of the age with ELISA and at 10 days of the age with AGP except one, but none of them showed from 1 day of the age. This report came to conclusions that both the protective maternal antibody titers and the antigen positive rates were significant until at the 10 days of the age.

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Production of Polyclonal Antibody against Grapevine fanleaf virus Movement Protein Expressed in Escherichia coli

  • Koolivand, Davoud;Bashir, Nemat Sokhandan;Behjatnia, Seyed Aliakbar;Joozani, Raziallah Jafari
    • The Plant Pathology Journal
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    • v.32 no.5
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    • pp.452-459
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    • 2016
  • The genomic region of Grapevine fanleaf virus (GFLV) encoding the movement protein (MP) was cloned into pET21a and transformed into Escherichia coli strain BL21 (DE3) to express the protein. Induction was made with a wide range of isopropyl-${\beta}$-D-thiogalactopyranoside (IPTG) concentrations (1, 1.5, and 2 mM) each for duration of 4, 6, or 16 h. However, the highest expression level was achieved with 1 mM IPTG for 4 h. Identity of the expressed protein was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting. The expressed 41 kDa protein was purified under denaturing condition by affinity chromatography, reconfirmed by Western blotting and plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) before being used as a recombinant antigen to raise polyclonal antibodies in rabbits. Purified anti-GFLV MP immunoglobulines (IgGs) and conjugated IgGs detected the expressed MP and GFLV virions in infected grapevines when used in PTA-ELISA, double antibody sandwich-ELISA, and Western blotting. This is the first report on the production of anti-GFLV MP polyclonal antibodies and application for the virus detection.

Studies on the Characteristics of Anti-Zone Antibody I. Identification of Anti-Zona Antibody by Indirect ELISA and Indirect Immunofluorescence Test (항투명대 항체의 특성에 관한 연구 I. 간접효소면역분석법과 간접면역형광분석법에 의한 항투명대 항체의 확인)

  • 김은영;유영춘;박세필;정형민;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.107-114
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    • 1990
  • These experiments were undertaken as a basic study to develop immunocontraceptive vaccine and to understand the role of zona pellucidae in early fertilization process by identifying the monoclonal and polyclonal antibody to porcine zona pellucidae and polyclonal antibody to mouse zona pellucidae by indirect ELISA and indirect immunofluorescence test. The results obtained in these experiments were summarized as follows : 1. The titer of the antibodies to zona was determined by indirect ELISA using solubilized porcine zona coated plates. Both monoclonal and rabbit polyclonal antibodies showed very high titers ; O.D at 1 : 12,800 dilution of antibodies was still significantly higher than that of non-immunized control serum. Rabbit anti-mouse zona pellucidae sera also reacted with porcine zona pellucidae. 2. By indirect immunofluorescence test strong fluorescences were observed on the egg treated with homologous and heterologous rabbit polyclonal antibodies and FITC lablled 2nd antibodies and found to crossreact strongly with the eggs from the pig and mouse. While weaken fluorescences were observed on the eggs treated with monoclonal antibodies.

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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.