• 제목/요약/키워드: ELISA method

검색결과 591건 처리시간 0.029초

ELISA 방법을 이용한 요중 아플라톡신 M1 측정 (Application of Competitive ELISA Method for Estimation of Urinary Aflatoxin M1 Level)

  • 김용대;김헌
    • 생명과학회지
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    • 제23권2호
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    • pp.306-310
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    • 2013
  • 본 연구는 요중 아플라톡신 M1 (AFM1)의 농도를 측정할 수 있는 competitive ELISA 방법의 특성을 HPLC-fluorescence detector (HPLC-FLD) 방법과 비교하여 평가하였다. ELISA 방법에서의 AFM1의 회수율은 105% (73-124%)였고 측정의 변이계수는 6.85%로 나타났다. ELISA 방법에서의 검출한계와 정량한계는 각각 0.20 pg/ml과 0.62 pg/ml로 조사되었으며, 두 방법을 이용하여 측정한 요중 AFM1 농도는 상관계수 0.96의 매우 높은 상관성이 있는 것으로 확인되었다(p<0.01). 그러나, 이렇게 높은 상관성에도 불구하고, ELISA 방법을 이용한 요중 AFM1의 농도는 HPLC-FLD 방법으로 측정한 값에 비해 상대적으로 높게 나타나는 경향을 보여 ELISA를 이용한 방법이 단시간에 많은 시료를 분석할 수 있는 장점은 있으나 그 결과는 HPLC-FLD 방법을 이용해서 얻은 회귀식을 이용하여 보정을 한 후 제시할 필요가 있는 것으로 판단된다.

Quantitative Analysis of Phosphinothricin-N-acetyltransferase in Genetically Modified Herbicide Tolerant Pepper by an Enzyme-Linked Immunosorbent Assay

  • Shim, Youn-Young;Shin, Weon-Sun;Moon, Gi-Seong;Kim, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제17권4호
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    • pp.681-684
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    • 2007
  • An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE3l-bar) and efficiently purified by $Ni^{2+}$ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: $0.01{\mu}g/ml$) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [$4.9{\pm}0.4{\mu}g/g$ of sample (n=6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.

곡류중 Ochratoxin A의 검색을 위한 면역분석법에 관한 연구 (Study on the Analysis Method of Ochratoxin A in Cereals by ELISA Method)

  • 김동술;정덕화;이용욱
    • 한국환경보건학회지
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    • 제20권3호
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    • pp.54-60
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    • 1994
  • We established ELISA method which is more rapid and safe than conventional analytical method, and then analyzed concentration of ochratoxin A in the rices by ELISA method. The results were as follows: 1. Since ochratoxin A molecular does not contain the group for coupling reaction, it was first proposed to have the function of an antigen. As a result of conjugation of ochratoxin A-BSA derivatives, one molecular of BSA was conjugated with 13 moleculars of ochratoxin A. 2. On the basis of established ELISA to apply for immuno analytical method of ochratoxin A, the minimal level of detection by ELISA method was at 0.5 ppb. 3. Rices (42) were collected from homes of Kyoungnam districts through November 1992 to December 1992, as a result of analysis, two of these were positive. Rice samples of R-1 and R-9 represented ochratoxin A levels of 6.0 ng/g and 10 ng/g, respectively.

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Establishment and application of a solid-phase blocking ELISA method for detection of antibodies against classical swine fever virus

  • Cao, Yuying;Yuan, Li;Yang, Shunli;Shang, Youjun;Yang, Bin;Jing, Zhizhong;Guo, Huichen;Yin, Shuanghui
    • Journal of Veterinary Science
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    • 제23권5호
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    • pp.32.1-32.11
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    • 2022
  • Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases. The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions: The spbELISA has practical applications in assessing the vaccination status of large pig herds.

유아용 조제분유 내 Cronobacter muytjensii 검지를 위한 간접 비경합 면역분석법의 개발 (Development of an Indirect Non-Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Cronobacter muytjensii in Infant Formula Powder)

  • 송신지에;김명희
    • 한국식품영양학회지
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    • 제26권4호
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    • pp.936-944
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    • 2013
  • Cronobacter muytjensii는 유아용 조제분유(IFP)의 잠재적 위험요인으로 중요한 식품 기인성 병원균이다. 이 연구에서는 C. muytjensii 검지를 위한 특이적 면역글로불린 G(IgG)를 개발하고, 이 anti-C. muytjensii IgG를 이용하여 간접 비경쟁 효소면역측정법(INC-ELISA)을 개발하였다. 그 결과, 새롭게 개발한 INC-ELISA 방법은 C. muytjensii에 매우 민감하고, 순수배양 시 $6.5{\times}10^3CFU/ml$의 검출한계와 유아용 조제분유에서 1 cell/25 g의 검출한계를 나타내었다. INC-ELISA 방법은 또한 C. muytjensii에 탁월한 특이성을 보이고, Cronobacter 속 외 11종의 다른 식품 기인성 병원균 계통과의 교차반응을 보이지 않았다. 이러한 결과는, 개발된 INC-ELISA 방법이 C. muytjensii에 매우 민감하고 효율적이며, 신속하고 용이한 검출을 위한 진단 키트 개발에 적용할 수 있음을 시사한다.

ELISA와 RT-PCR에 의한 국내재배난에서 심비디움 모자이크 바이러스와 오돈토글로섬 윤문 바이러스이 검정 (Detection of Cymbidium Mosaic Virus and Odontoglosum Ringspot Virus by ELISA and RT-PCR from Cultivated Orchids in Korea)

  • 박원목;심걸보;김수중;류기현
    • 한국식물병리학회지
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    • 제14권2호
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    • pp.130-135
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    • 1998
  • This study was carried out to detect cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamovirus (ORSV) in cultivated orchid plants in Korea. The standard double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) were carried out for detection of the viruses in the collected orchid samples. ELISA was suitable for massive-scale diagnostic method for virus detection in orchids. RT-PCR was rapid, time-saving and reliable detective method, and detection limit data showed that RT-PCR was 103 times more sensitive than ELISA. Of the 321 individual orchids representing 5 orchids genera tested by the ELISA, CymMV and ORSV were detected in 15.6% and 22.4%, and mixed infection of the both viruses with 4.9%, respectively. Of the Cymbidium plants tested, cultivated plants showed 52.5% virus infection rate with either CymMV or ORSV and both viruses.

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조직인공소화법과 ELISA를 이용한 국내 출하돈의 선모충(Trichinella spiralis) 감염실태 조사 (The survey of Trichinella spiralis infection in finishing pigs using the pepsin-digestion method and ELISA in Korea)

  • 서흔수;우계형;윤희정
    • 대한수의학회지
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    • 제44권2호
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    • pp.269-277
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    • 2004
  • Trichinella spiralis is one of the important zoonotic parasites with a wide variety of vertebrates hosts in nature. The purpose of this study were to analyze ESP(Excretory-Secretory Protein) antigen, to evaluate ELISA for the serological diagnosis of Trichinosis, and to survey T. spiralis infection in finishing pigs using the pepsin digestion method and ELISA in Korea. In the analysis of ESP antigen by SDS-PAGE and Western blot, 4 major bands (70, 55, 52.6, and 49 kDa) were revealed from the ESP antigen. Predilection sites of T. spiralis were the diaphragm, the tongue, masseter muscles, intercostal muscle, and hindlimb in orders in the experimentally infected rats. Sera from 581 swine were tested by ELISA with ESP antigen. The 54 (9.3%) sera were suspected as positive reactors, however, these 54 sera were determined as false positives by the use of Western blotting. This study demonstrated that the ELISA was not suitable for the examination of T. spiralis in pork. The diaphragm muscle samples of 251 finishing pigs were tested by the method of pepsin-digestion for the presence of Trichinella larvae, however, T. spiralis was not detected from the samples. We could not find out T. spiralis infection in pig in Korea pork.

만성 기침 환자에서 혈청 클라미디아 항체에 대한 ELISA와 microimmunofluorescence 검사의 비교 (Enzyme-linked Immunosorbent Assays for Antibodies against Chlamydia Pneumoniae Compared with Microimmunofluorescence Test with Patients with Chronic Cough)

  • 이희영;김우진
    • Tuberculosis and Respiratory Diseases
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    • 제59권1호
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    • pp.47-52
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    • 2005
  • 연구배경 : 클라미디아 폐렴균은 임상적으로 중요한 균으로 혈청검사가 진단에 중요하다. 현재 이용되는 micro-immunofluorescence법은 주관적이고 시간이 많이 소요되므로 좀더 객관적으로 진단할 수 있는 ELISA법을 이용하여 만성 기침을 주소로 내원한 환자들의 혈청을 대상으로 검사하고 표준방법과 결과를 비교하였다. 방 법 : 2003년 8월부터 2004년 7월까지 강원대학교병원을 방문한 35명의 성인 환자에서 얻은 혈청을 ELISA법과 micoimmunofluorescence법으로 시행하였다. 비인두 스왑에서 PCR을 시행하였다. 두 방법의 비교는 비모수 상관계수 분석(Spearman)을 하였다. 결 과 : ELISA 검사결과 IgG는 민감도 84.0%, 특이도 60.0%, IgA는 민감도 84.0%, 특이도60.0%, IgM은 민감도 40.0%, 특이도 96.7%의 결과를 보였다. PCR은 3명의 환자에서 양성이었다. 결 론 : ELISA법이 많은 환자들을 대상으로 하는 역학적 연구에서는 좀더 유용하게 이용될 수 있을 것으로 사료된다. 임상적으로 적용되기 위해서는 좀더 연구가 필요할 것으로 사료된다.

Enzyme-linked Immunosorbent Assay(ELISA)를 이용한 혈청 및 원유 중의 Mycobacterium bovis 항체 검출에 관한 연구 (Studies on Enzyme-linked Immunosorbent Assay(ELISA) for Detection of Antibody to Mycobacterium bovis in Serum and Milk)

  • 심항섭;국정희;박병옥;김성열;박유순
    • 한국동물위생학회지
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    • 제20권2호
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    • pp.133-142
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    • 1997
  • In order to supplement a diagnostic method for detection of infectious cattle to bovine tuberculosis, performed ELISA for detection of antibody to if bovis in serum and milk. The diagnostic efficacy of the established ELISA was compared with test of the tuberculin skin test for bovine tuberculosis. The positive corresponding rate of serum ELISA and tuberculin skin test showed 84.3%, milk ELISA and tuberculin skin test showed 75.0%, milk ELISA and serum ELISA showed 75.0% respectively. Comparison of the serum and milk to tuberculin antibody concentration in tuberculin positive cattle, the milk contained 1/100-1/150 concentration compared serum tuberculin concentration. The established ELISA was considered efficient for detection of antibodies to M bovis in serum and milk.

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간흡충증에 있어서 항체검출을 위한 Enzyme-linked Immunosorbent Assay와 Thin Layer Immunoassay의 비교 (Comparison of TIA with ELISA for circulating antibody detection in clonorchiasis)

  • 이용기;유재숙이근태정경일
    • Parasites, Hosts and Diseases
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    • 제21권2호
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    • pp.265-269
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    • 1983
  • 간흡충증 환자 혈청에서 Thin layer immunoassay에 의하여 항체를 정량적으로 측정하여 감수성과 특이성을 알아보고, 효소표식면역 법(ELISA)에 의한 성적과 비교하였다. 냉동 건조한 간흡충 성충의 생리식염수 추출물을 항원으로 사용하였고, TIA는 Elwing등(1976)의 방법으로, 효소표식면 역법은 Veiler등(1974)의 방법에 의해 실시하였다. 1. 간홉충중 환자 60명의 혈청에서 TIA zone 크기의 평균은 4.14mm였고, 3mm 이상을 양성으로 판정하였을 때 감수성은 100%, 특이성은 61.1%였다. 2. 효소표식면역 법에 의하면 ELISA치 0.8이상을 양성으로 판정하였을 때 감수성은 88.3%, 특이성은 87.3%였다. 3. 혈청내 IgG 농도는 TIA zone 크기와 상관이 없었으나 좌소표식면역법에 의한 ELISA치와는 상관이 있어 그 계수는 0.69였다. 4. TIA zone 크기는 효소표식면역 법에 의한 ELISA치와 그 분포에 있어 상관이 없었다.

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