• Title/Summary/Keyword: 면역분석법

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식품내 진균독소분석을 위한 면역화학적 방법의 평가와 응용

  • Lee, Hyang-Beom;Son, Dong-Hwa
    • Bulletin of Food Technology
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    • v.10 no.1
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    • pp.115-120
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    • 1997
  • 면역분석법(immunoassay)은 aflatoxin류, deoxynivalenol(DON), zearalenone(ZEA), fumonisin류 등을 포함하는 다양한 진균독소분석에 적용되고 있다. 이들 분석법을 이용하여 여러 종류의 matrix내에 존재하는 물질들을 분석할 수 있다. 그리고 많은 시료를 빨리 분석해야 하는 경우에 이 방법이 적용되고 있다. 상업적인 면역화학적 분석키트들은 흔히 검출감도(sensitivity), 특이성(specificity), 재현성(reproducibility), 비용(cost), 안정성(stability), 편리성(ease of use)등의 관점에서 평가된다. 실험실적으로 품질보증을 위하여 표준곡선과 양성대조구 등에 대한 사전 검토가 필수적이다. 또한 높은 농도의 분석물의 경우에는 matrix에서 기인한 간섭때문이 아닌지를 구분하는 것도 중요하다. 본문에서 곡류와 곡류가공품 및 특정의 감시대상물 질내 aflatoxin류, DON, ZEA 그리고 fumonisin류와 같은 진균독소의 분석에 면역화학적 방법을 사용하여 얻어진 공동연구의 결과에 대한 해석기준이 제시된다. 또한 yes/no로 구분해야 하는 정성시험의 결과해석에 필요한 기준이 제시된다. 진균독소의 면역분석법을 평가하기 위한 몇몇기준들과 그 방법들을 감시도구로 사용하는 예는, 식품중의 잔류농약을 검출하기 위한 접근방법과 유사한 모델로서 활용될 수 있을 것이다.

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Further Systematic Studies on Cornus and Relatives by Immunoelectrophoresis (면역학적 전기영동에 의한 층층나무속과 그 근연군의 계통학적 추가연구)

  • Lee, Yoo-Sung
    • Journal of Plant Biology
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    • v.28 no.2
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    • pp.95-104
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    • 1985
  • Additional serological data from radial immunodiffusion and conventional immunoelectrophoresis with combination of presaturation of antibody were obtained for further interpretation of Cornus (and relatives) systematics. Pollen proteins were compared by qualitative and quantitative means. Cornus drummondi and C. stolonifera are very similar to C. florida, and C. amomum and C. recemosa are less similar. Nyssa constitutes the next distinct and most similar family, Nyssaceae to the Cornaceae. The serological affinities indicate that Corokia and Griselinia should not be included within the Cornaceae. Their taxonomic treatment to elevate to the family level is now awaiting until more data are accumulated.

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Comparison of chemiluminescence immunoassay (CIA) and immunochromatography assay(ICA) for Detecting HBsAg and Anti-HBs (B형 간염항원 및 항체 검사를 위한 화학발광면역검사법(CIA)과 면역크로마토그래피법(ICA)의 비교분석)

  • Kim, Hye-Jeong;Kim, Eun-Hee
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.7
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    • pp.3419-3424
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    • 2013
  • Diverse immunoassays including a chemiluminescent immunoassay (CIA) are used to detect hepatitis B surface antigen (HBsAg) and antibody (anti-HBs). Recently, an increasing number of institutions have been utilizing an immunochromatography assay (ICA), which is easy to use. In this study, We evaluated ICA kits for the rapid detection of HBsAg and anti-HBs by comparing them with a CIA. A total of 120 serum hospital samples, were collected for the whole month, were assayed using ICA kit. The Concordance rate, sensitivity, specificity, positive predictive value and negative predictive value of the ICA for HBsAg based on CIA results were 97%, 97%, 100%, 100%, and 96.8%, respectively. The diagnostic performances of the ICA for Anti-HBs were 90%, 90%, 93.3%, 93.1%, and 90.3%, respectively. The ICA kit failed to detect HBsAg and anti-HBs in low reactive samples. The ICA kits for the rapid detection of HBsAg might be recommended for interpreted with caution and dual analysis in the clinical laboratory.

Analysis of Salmonella Species from Eggs Using Immunoliposomes and Comparison with a Commercial Test Kit (면역리포좀을 이용한 계란에서의 살모넬라 분석과 시판 간이키트와의 비교)

  • Shin, Weon-Sun;Kim, Yoon-Sook;Lee, Jun-Soo;Kim, Myung-Hee
    • Food Science of Animal Resources
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    • v.29 no.4
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    • pp.533-538
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    • 2009
  • To suggest an improved diagnostic method for Salmonella spp., analyses were conducted with immunoliposomes and compared with the results from a commercial test kit. One sample out of 36 samples of eggshell was Salmonella-positive via immunoliposomes. In the case of the use of the commercial test kit, six samples out of 36 samples were Salmonellapositive. These Salmonella-positive samples were subjected to biochemical identification tests that confirmed that they were Salmonella-negative. As for the egg content samples, they were Salmonella-negative in both analyses with immunoliposomes and the commercial test kit. The Salmonella analysis with immunoliposomes reduced detection time, by 24 h compared to the commercial test kit. Bacteria, including Acinetobacter baumanni, Chryseomonas luteola, Enterobacter cloacae, Escherichia coli, Escherichia hermannii, Klebsiella pneumonia, Pantoea spp., and Pasteurella pneumotropica, were isolated from the eggshells. Other than Acinetobacter baumanni and Pasteurella pneumotropica most of the isolates were known to frequently appear during egg production processing.

Production of Monoclonal Antibodies against Vibrio parahaemolyticus and Development of High Sensitive Immuno-Selective Filtration Method (Vibrio parahaemolyticus에 대한 단클론성 항체 개발과 고감도 면역선택여과법의 개발)

  • Kim, Jeong-Sook;Choi, Young-Dong;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.21-27
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    • 2016
  • The objectives of this study are to produce monoclonal antibodies (MAbs) against Vibrio parahaemolyticus and to develop an immuno-selective filtration (ISF) method for the rapid and sensitive detection of V. parahaemolyticus. The characterization of the MAb produced from HKVP 4H9-9 hybridoma cell was validated by enzyme-linked immunosorbent assay (ELISA) and western blot. The produced MAb was specific to V. parahaemolyticus and showed weak cross-reaction to V. alginolyticus, V. vulnificus and Staphylococcus aureus. After optimization of the method, $5{\times}10^1cell/mL$ of V. parahaemolyticus in a pure culture could be detectable. Although weak cross-reactivity to V. vulnificus, V. alginolyticus and Staphylococcus aureus was observed, the ISF was confirmed to be highly specific to V. parahaemolyticus. Especially, the ISF showed the most sensitivity compared to the immunoassays currently reported is easier to perform and quicker than ID-ELISA.

Production of Polyclonal Antibody against $\alpha$-Fetoprotein and Polyclonal Antibody-Based Competitive Enzyme-Linked Immunosorbent Assay for $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein (AFP)에 대한 폴리클로날 항체의 생산 및 $\alpha$-fetoprotein 측정용 효소면역분석법 (competitive ELISA)의 개발)

  • Michung Yoon
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.115-123
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    • 1997
  • $\alpha$-Fetoprotein (AFP) has been a useful marker in screening and/or monitoring patients with hepatocellular carcinoma, gonadal germ cell tumor, gastric carcinoma and neural tube defects. In the present study, it was attempted to produce anti-human AFP polyclonal antibodies and to develop a competitive enzyme-linked immunosorbent assay (ELISA) for the measurement of AFP in human plasma and amniotic fluid. AFP was isolated from amniotic fluid using an isolation procedure consisting of affinity chromatography and preparative polyacrylamide gel electrophoresis. The antibody directed against AFP was raised in rabbits. Double immunodiffusion and Western blotting methods showed that the antiserum was highly specific, reacting with only AFP-containing samples. Standard curve was obtained by using purified AFP and specific antiserum. The assay sensitivity was 5ng/ml and the working range was 5~l,000ng/ml. The within-assay and between-assay coefficient of variance (CV) was 4.5% and 8.5%, respectively. These results indicate that the assay is valuable for the measurement of AFP and found to be simple, reproducible, and accurate.

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유청 단백질에서 유도되는 생리활성 펩타이드에 관한 연구

  • Yun, Seung-Seop
    • 한국유가공학회:학술대회논문집
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    • 1996.11a
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    • pp.18-29
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    • 1996
  • 1. CWPC중의 새로운 생리활성물질의 검색 Mouse 임파세포의 증식효과를 지표로 하는 면역기능을 검토하여 CWPC중의 면역 부활작용을 갖는 새로운 성분의 검색을 실시하였다. CWPC를 여러 가지 분획법으로 분획하여 mouse 임파세포의 증식효과를 지표로 면역 활성성분을 검색하였다. 그 결과 gel filtration, 음이온교환법을 사용하여 분획한 당을 다량 포함한 부분에 강한 면역부활담당세포에 대하여 증식활성을 나타내는 물질을 발견하였다. 이 물질은 SDS-PAGE상에서 분자량이 약 16kDa에 위치하여 Ca, P 및 당쇄를 포함한 물질이며, 이것을 GPP로 하였다. GPP에는 우유케이신의 trypsin분해물이며 Ca와 무기인을 풍부하게 포함하는 ${\beta}$-CPP와 유사한 phosphoserin 영역을 갖는 성분과 갖지 않는 성분의 2종류가 존재하며, 각각의 면역 부활활성이 인정되었다. 각 성분의 아미노산 분석, 당 분석의 결과에서 지금까지 보고된 우유중의 면역 담당세포에 대한 증식활성을 갖는 물질과는 상이한 성분인 것으로 밝혀졌다. 더욱이 이 활성물질 (GPP)은 PP cell에서도 동등한 활성이 있는 것으로 판단되었다. 이러한 결과를 종합하여 보면 CWPC중에서 지금까지 알려지지 않았던 새로운 면역 부활물질이 존재하며, 그 성분에는 CPP와 유사한 phophoserine 영역이 존재하는 성분이 포함되어 있고, N-글리코실 결합의 당쇄가 존재하는 것으로 시사되었다. 이 성분은 전신면역의 지표인 비장세포에 대해서만이 아니고, 장관면역계에 중요한 역할을 담당하는 PP cell에서도 활성이 있는 것으로 보아 전신 및 국부적인 면역기능의 부활성분으로서 응용의 가능성이 시사되었다. 2. GPP의 면역담당세포에 대한 증식활성의 메카니즘의 검토 CWPC중의 GPP의 면역담당세포증식활성의 메카니즘을 해명하기 위해 먼저 이 성분중의 어느 부분이 활성에 관여하는지를 pronase 분해 및 phophoserine 영역을 인식하는 항체를 사용하여 검토하였다. 그 결과 pronase 분해처리에서도 활성의 감소를 나타내지 않았으므로 이러한 활성에는 당이 필수 불가결하다는 점이 시사되었다. 또한 phosphoserine 영역을 인식하는 항체에 의해서도 활성은 감소하지 않는 것으로 보아 phosphoserine 영역이 세포증식활성에 관여하지 않는 것으로 판단되었다. 또한 분획한 면역담당세포에 대한 증식활성을 측정하는 것으로 이 성분의 표적면역담당세포를 동정하여, B세포에 대해서만 특이적으로 증식활성을 나타내는 것으로 밝혀졌다.

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Immunoelectrophoretic analysis of major component proteins In cystic fluid of Taenia solium metacestodes (면역전기영동법에 의한 유구낭미충 낭액의 구성 단백질 분석)

  • Yoon Kong;Seung-Yull Cho;Suk-Il Kim;Shin-Yong Kang
    • Parasites, Hosts and Diseases
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    • v.30 no.3
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    • pp.209-218
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    • 1992
  • When cystic fluid of Taenia solium metacestodes (CF) was filtrated through Sephacryl S-300 Superfine, major proteins were in fractions III add IV Major protein in fraction III was Band C protein of 150 kDa and that in fraction IV was Band N protein (Choi et of., 1990). When CF was electrophoresed in 0.9% agarose gel and reacted with anti-CF rabbit serum (RACF), two main bands, a long outer and a short inner band, were precipitated, together with 8 minor bands. RACF reacted with fraction III forming the long outer band whereas RACF formed the short infer band with fraction IV in immunoelectrophoresis (IEP) The long outer precipitin band of CF fraction III was similar to antigen B in hydatid fluid (HF) of Oriol et at. (1971), while the short inner band of CF fraction IV was similar to HF antigen 5 of Caption et at. (1967) . When HF was reacted with RACF, the short inner band was immunoprecipitated without forming the long outer band. Common antigenicity between CF and HF seemed to exist in fraction IV rather than in fraction III of CF. Patient sera of neurocysticercosis reacted more frequently with fraction III than with fraction IV.

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Electrochemical Immunoassay based on the Dopamine-antigen Conjugate for Detecting Hippuric Acid (항원인 마뇨산에 결합된 도파민을 이용한 전기화학적 면역 분석법)

  • Choi, Young-Bong;Jeon, Won-Yong;Kim, Hyug-Han
    • Journal of the Korean Electrochemical Society
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    • v.17 no.3
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    • pp.172-178
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    • 2014
  • In this work, we describe an electrochemical immunosensor for simple, fast and quantitative detection of a urinary hippuric acid (HA). Urinary HA, of molecular weight 180 DA, is one of the major metabolites and biological indicators in toluene-exposed humans. Simple and ubiquitous monitoring of exposure to toluene is very important in occupational health care. We propose the electrochemical immunoassay based on the dopamine-antigen conjugate for detecting hippuric acid. Our electrochemical immunoassay system employs a conjugate of dopamine (DA) as an electrochemical active molecule and hippuric acid (HA) as an antigen. As an electrochemical aspect, dopamine (DA) containing two hydroxyl group can show excellent redox signal. Also, dopamine-tethered hippuric acid (DA-HA) shows the reversible redox signal in the immunoassay. The competition between HA and DA-HA generated electric signals proportional to HA concentration. The electrochemical immunoassay was performed with DA-HA on the screen printed carbon electrodes (SPCEs), and then applies the mixture antigen (HA) and HA-antibody. The electrical signals were proportional to HA in the range of 0.010~2.500 mg/mL which is enough range to be used for the point-of-care.