• Title/Summary/Keyword: ELISA method

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Application of Immunoassay for the Detection of Gamma-Irradiated Shrimp (감마선 조사된 새우의 검지를 위한 면역분석법의 적용)

  • 이주운;육홍선;조경환;차보숙;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.4
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    • pp.600-604
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    • 2001
  • Immunoassay was used to study the detection method of irradiated shrimp. Sandwich ELISA was formatted with monoclonal antibody (Ab) (M-IgG) and polyclonal Abs (P-IgG) and polyclonal Abs (P-IgG) individually produced against brown shrimp tropomyosin (TPM) as an antigen. When M-IgG was used as a coating Ab to capture TPM, and P-IgG were used as reaction Ab against captured TPM could be detected in the range of 12.5 to 50 $\mu\textrm{g}$/mL. Detected concentrations of TPM from irradiated shrimp decreased dose-dependently, and the concentration of Ag by combination of irradiation with heating or freezing treatments also decreased. This results suggests the possibility for Sandwich ELISA, one of immunological analyses, to be applied for detecting irradiated shrimp.

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Detection of Xanthomonas axonopodis pv. citri on Citrus Fruits Using Enzyme-Linked Immunosorbent Assay

  • Jin, Kyoung-Sik;Kang, Ik-Beom;Ko, Kyoung-Il;Lee, Eun-Seob;Heo, Jong-Young;Kang, Young-Kil;Kim, Byung-Ki
    • The Plant Pathology Journal
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    • v.17 no.1
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    • pp.62-66
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    • 2001
  • Detection of Xanthomonas axonopodis pv. citri (Xac) on citrus fruits for exporting is usually made by bacteriophage test (BPT) to demonstrate the pathogen-free status. BPT has rather time-consuming and complicate procedures for dealing with massive samples to be inspected. In this study, enzyme-linked immunosorbent assay (ELISA) was applied to detect Xac on fruits, and compared with BPT. In ELISA, positive reactions occurred in the bacterial densities of $3\times10^5$ cells/ml or more. To detect the bacterial infection on citrus fruits with a density of lower than $3\times10^5$ cells/ml, the bacterial suspensions were mixed with fruit rinse water and incubated in broth medium. Ordinary peptone sucrose broth (PSB) was not a proper medium for increasing Xac density specifically enough to be detect by ELISA. On the other hand, modified PSB (MPSP) amended with Fe-EDTA (0.25 g/$\ell$) and 2.5% potato-dextrose broth sufficed to differentiate uninfected and infected citrus fruits by ELISA after 24 h incubation of the fruit rinse water. Using various citrus samples from infected and uninfected fields, efficiencies in detecting Xac on fruits were compared between ELISA and BPT. For infected fruits samples, ELISA detected Xac by 100%, while BPT by about 44%, indicating that the detection efficiency was improved by 23.5% by ELISA, compared to BPT. In addition, ELISA has simpler procedures for testing and is less time-consuming than BPT, suggesting that ELISA may be accurate and simple method to detect Xac on citrus fruits.

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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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A Simple and Fast Microplate Method for Analysis of Carotenoids Content in Chili Pepper (Capsicum annuum L.) (고추의 카로티노이드 함량 분석을 위한 신속·간편 마이크로플레이트법)

  • Hong, Sun Chul;Han, Jung-Heon;Lee, Jundae;Ahn, Yul Kyun;Yang, Eun-Young;Chae, Soo Young;Kim, Su;Yoon, Jae Bok
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.807-812
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    • 2013
  • In this study, we aimed to develop a simple analysis method for measuring the carotenoids content of pepper powder. A 96-wells polystyrene microplate and an ELISA reader were used for analysis. Although ELISA reader with 450 nm filter was applicable to measure carotenoid contents, the surface of microplates were degenerated by acetone used for carotenoids extraction. However, ten-folded dilute of the color extract with methanol did not affect the surface of polystyrene microplate and components of the color extract could be successfully measured by a ELISA reader, showing a high corelation with ASTA-20.1 method. In addition, this method uses 10 fold less acetone than ASTA-20.1 method resulting less acetone waste. The microplate method using ELISA reader has potential power for analyzing a large number of samples which may be very useful to the practical breeding program for high-colored peppers.

Serodiagnosis of Extraintestinal Amebiasis: Retrospective Evaluation of the Diagnostic Performance of the Bordier® ELISA Kit

  • Beyls, Nicolas;Cognet, Odile;Stahl, Jean-Paul;Rogeaux, Olivier;Pelloux, Herve
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.71-74
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    • 2018
  • Soluble antigens from an axenic culture of Entamoeba histolytica were used to develop a commercial ELISA kit to quantify anti-E. histolytica antibodies in sera of patients with extraintestinal amebiasis in non-endemic settings. The diagnostic specificity and sensitivity of the test were assessed retrospectively using 131 human serum samples with amoebic serologic status available. They were selected according to their results in immunofluorescence (IFAT) and were separated in 2 sample categories: 64 sera with positive results by IFAT and 67 with negative results by IFAT. The sensitivity and specificity of the ELISA kit were assessed at 95.0% and 94.0% compared to the IFAT. The test can be useful to exclude a potential diagnosis of amebiasis and could be used as a screening method since ELISA is an automated technique.

A Dipstick-Type Enzyme-Linked Immunosorbent Assay for the Detection of the Insecticide Fenitrothion in Food Samples

  • Cho, Young-Ae;Shim, Jee-Youn;Lee, Yong-Tae;Lee, Hye-Sung
    • Food Science and Biotechnology
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    • v.15 no.6
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    • pp.990-992
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    • 2006
  • In a previous study, we obtained polyclonal antibodies against the organophosphorus insecticide fenitrothion and developed an enzyme-linked immunosorbent assay (ELISA) for this pesticide. Using these antibodies and an enzyme tracer, a direct competitive ELISA method specific for fenitrothion using a dipstick format was developed. Dipstick ELISA using antibodies to fenitrothion immobilized on an Immunodyne membrane allowed the quick visual detection of fenitrothion at concentrations above $10\;{\mu}g/L$. The $IC_{50}$ value of dipstick ELISA using reflectance detection was $27\;{\mu}g/L$ with a detection limit of $2\;{\mu}g/L$. The recovery of fenitrothion from spiked lettuce and rice samples using the dipstick ELISA ranged from 87-107%.

Detection of IgG and IgM antibodies with ELISA technique in human trichomoniasis (질트리코모나스 환자에서 효소표식 면역검사법을 이용한 혈청 내 항-질트리코모나스 IgG 및 IgM 항체가의 측정)

  • 이미리;신명헌
    • Parasites, Hosts and Diseases
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    • v.28 no.1
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    • pp.25-30
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    • 1990
  • The direct wet mount examination of vaginal. secretion, widely applied for the diagnosis of Trichcmonas vaginalis infection in woman patients, is rapi4 and economical, however, the sensitivity of this technique is not so high. In this study enzyme-linked immunosorbent assay (ELISA) was employed for the detection of serum anti-T. vaginalis IgG and IsM antibodies from 30 vaginal trichomoniasis patients and 30 non-infected healthy persons. The results were as follows: 1. Serum ELISA-IgG value was $0.37{\pm}0.134(Mean{\pm}S.D.)$ in vaginal trichomoniasis patients and $0.21{\pm}0.054$ in healthy controls(P<0.005), and the sensitivity and specificity of ELISA for serum IgG antibody were 70.0% and 96.7%, respectively. 2. Serum ELISA-IgM value was $0.33{\pm}0.177 (Mean{\pm}S.D.)$ in vaginal trichomonlasls patients and $0.11{\pm}0.051$ in healthy controls (p<0.005), and the sensitivity and specificity of ELISA for serum IsM antibody were 70.0% and 96.7%, respectively. 3, The ELISA-IgG values showed a significant correlation with ELISA-IgM values(r=0.77, p<0.005) , With above results, it is assumed that ELISA is a reliable method for the diagnosis of T vaginalis infection and simultaneous measurement of serum IgG and IgM with this technique is recommended.

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Development and Evaluation of a Competitive Enzyme Immunoassay for the Detection of Antibodies to Treponema pallidum (경쟁적 효소면역 측정법을 이용한 Treponema pallidum 항체 진단시약의 개발 및 평가)

  • 김병문;이정환;정문섭;김승철;이미용;이성희;김원배
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.344-348
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    • 1999
  • A competitive enzyme-linked immunosorbent assay(ELISA) for the detection of antibodies to Treponema pallidum(T.pallidum) was developed and evaluated. T.apllidum lysate was immobilized on the surface of microplate wells and horseradish peroxidase labeled human anti-T.pallidum lysate was immobilized on the surface of microplate wells and horseradish peroxidase labeled human anti-T.pallidum was prepared and used as a tracer. The performance of the competitive ELISA was evaluated by using different specimens. The competitive ELISA showed a sensitivity of 100% in a performance panel consisting of serum and plasma with anti-T.pallidum reactivity ranging from negative to strong positive by FTA-ABS test system and 120 plasma samples positive by TPHA. The specificity of the competitive ELISA was 100% in 1,200 plasma samples collected from healthy seronegative blood donors. These results suggest that the competitive ELISA provides an excellent assay method for the detection of antibodies to T.pallidum, and may be particularly useful for serological blood screening of syphilis.

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Studies on serological tests for pullorum disease (추백리의 혈청학적 진단법에 관한 연구)

  • 김정태;심항섭;김태종;고태오;우종태;유기승;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.3
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    • pp.313-323
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    • 1998
  • In order to establish a sensitive and specific diagnostic method for detection of antibody to Salmonella pullorum, a enzyme-linked immunosorbent assay(ELISA) was designed and standardized. The diagnostic efficacy of the established ELISA was compared with that of the serum plate agglutination test and immunodiffusion test for pullorum disease. 1. The chicken hyperimmune sera to Salmonella pullorum, S gallinarum, S typhimurium and S typhi were shown the cross reaction to S pullorum antigen by serum plate agglutination test. 2. When compared the cross reaction titer of microplate agglutination test for chickens hyperimmune sera, it was found that the titer were 64 in S pullorum, 32 in S gallinarum, 4 in S typhimurium and 8 in S typhi, respectively. 3. When compared the specificity of various antigen(HA, EA, PA and SA) by the immunodiffusion test, the most suitable antigen was phenol-treated bactrium. 4. The optimal concentration of S pullorum antigen for ELISA was 1 : 160 dilution of bacterium. 5. The efficacy of the ELISA for detection of S pullorum antibody was compared with serum Plate agglutination test and immunodiffusion test in chickens infected with S pullorum. The antibody was first detected at 6 days after infection using three tests examined. The antibody was alldetected at 9 days by ELISA, at 12 days by serumplate agglutination test, at 15 days by immunodiffusion test.

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Screening of Ochratoxin A Producing Fungi from Greenhouse Horticulture (시설원예산물로부터 Ochratoxin A 생성 곰팡이의 검색)

  • Kang, Sung-Jo;Park, Bong-Jung;Lee, Jong-Ok;Kang, Jin-Soon;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1415-1419
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    • 1998
  • In order to evaluate the safety of greenhouse horticultures, a large number sample sources were collected, and the fungi of Aspergillus sp. and Penicillium sp. were isolated from them. Indirect competitive ELISA method and high performance liquid chromatography (HPLC) were applied to confirm the ochratoxin A producing abilities of isolated strains. One hundred ninety two sample sources including soil, pepper, strawberry and water mellon were collected for fungi isolation from western Gyeongnam, Andong and Gyeongbok. One hundred forty two strains of Aspergillus sp. and one hundred fifty three strains of Penicillium sp. were isolated respectively from them. The isolated fungi were tested for the production of ochratoxin A by ELISA. After culture of them on the modified sucrose low salt medium at $28^{\circ}C$ for 15 days, we found that five strains of Penicillium sp. produced ochratoxin A at the levels of $0.084{\sim}2.128\;{\mu}g/mL$. Among them, #129-2 strain isolated from water melon, showed the highest level of ochratoxin A as $2.128\;{\mu}g/mL$ broth. However, all of isolated Aspergillus sp. didn't produce ochratoxin A. When we compared the results of ELISA method with HPLC method, ochratoxin A production of each isolated strains showed very similar levels.

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