• Title/Summary/Keyword: 효소면역법

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Comparison of Conventional Culture Method, Enzyme Immune Method, and PCR for the Rapid Detection of Salmonella spp. in Pet Food (반려동물 사료의 Salmonella spp. 신속검출을 위한 증균배양법, 면역학적 검출법 및 종 특이 프라이머를 이용한 PCR 방법 비교)

  • Yun, Hyejeong;Cha, Sun Ho;Lee, Seung-Hwa;Jeong, Min-Hee;Na, Tae-Woong;Kim, Haejin;Cho, Hyunjeong;Hong, Seong-Hee
    • Journal of Food Hygiene and Safety
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    • v.37 no.1
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    • pp.15-20
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    • 2022
  • The purpose of this study was to compare the conventional culture method, enzyme immune method and the PCR method using species-specific primer in the analysis on the Salmonella spp. found in domestically distributed pet foods. For the study, Salmonella spp. were detected from 175 samples. From the conventional culture method and the PCR method, two samples (jerky and corn gluten) were determined as positive. Also, from the enzyme immune method, one sample (corn gluten) was test-positive. The study revealed that application of the PCR method with species-specific primer allows better distinguishment between the species of the strain collected from the samples than the conventional culture method and/or the enzyme immune method.

The Evaluation of Recornbinant Circumsporozoite Protein in Malaria Diagnosis (말라리아 진단시 재조합 Circumsporozoite 단백질의 유용성 평가)

  • 이형우;이종수;이원자;조신형;이호자
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.142-149
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    • 2000
  • It had been evaluated the recombinant Circumsporozoite(CS) protein of Plasmodium viva in serologic diagnosis of vivax malaria. Western blot was done to analyse the sera of malaria patients according to the days after onset. The sera which have the terms within 15 days were shown 43.8%(14/32) of positive rates and the sera over the 16 days were shown 94.4%(17/18) of positive rates. So the total positive rate was 62%(31/50). It was 22.6%(7/31) which was shown negative response in Western blot, even though they were shown positive response in Immuuofluorescent antibody test(1FAT) using whole blood stage antigens. The positive rate of non-epidemic area(Yechon-gun, Kyongsangbuk-do) was 10.7%(3/28), and epidemic area(Kangwha-gun, Inchon-shi) was 27.6%(13/47) in Western blot analysis using recombinant CS protein. In order to applicate the recombinant CS protein in seroepidemiological survey, blood samples of 422 inhabitants were collected who lived in malaria epidemic areas, Chosm-ri, Majeong-ri, Hyangyang-ri and Noejo-n in Paju-shi, Kyonggi-do. All of them were negative in microscopic examination and two(0.5%) of them were positive in Polymerase Chain Reaction. 42(10.0%) of them were seropositive in FAT using whole blood antigens and 71(16.8%) of them were seropositive in Enzyme-linked immunosorbent assay using recombinant CS protein. It was figured out the positive rates were much higher according to the distances of villages which were closed to the demilitalized zone(DMZ) in all kind of diagnostic methods, respectively.

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Measurement of the Affinity Constant of Monoclonal Antibody to Human Apolipoprotein A-I by ELISA (효소면역 분석법에 의한 아포지단백질 A-I 단일클론항체의 친화상수의 측정)

  • Mic Hung Yoon;Hyun Hee Lee
    • Biomedical Science Letters
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    • v.1 no.1
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    • pp.1-8
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    • 1995
  • The present study was undertaken to determine the dissociation constant (Kd)of monoclonal antibody to human apolipoprotein A-I (apo A-I) using enzyme-linked immunosorbent assay (ELISA). First the monoclonal antibody was incubated in solution with the antigen until the equilibrium was reached; then the free antibody which remains unsaturated at equilibrium was captured by binding to antigen on the microtiter plate and be measured by a classical indirect ELISA. The value of Kd determined from Scatchard plot was 0.625$\times$10^{-9}$ for purified antibody and 0.720$\times$10$^{-9}$ for unpurified antibody. This method was valuable for the measurement of true dissociation constant and found to be simple, reproducible, and accurate.

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Enzyme-Linked Immunosorbent Assay for Identification of Irradiated Eggs (효소면역 측정법에 의한 방사선 조사 계란의 검출)

  • 이경애;최윤정;양재승
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.6
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    • pp.1030-1034
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    • 2000
  • The ovalbumin, a most sensitive egg white protein to irradiation was purified from irradiated hen's eggs. Eggs were irradiated in their shells to 0~7 kGy. To investigate for a practical use in identifying of irradiated eggs, competitive ELISA using ovalbumin was peformed. The binding activity of ovalbumin to anti-ovalbumin IgG was reduced in a dose-dependent manner by irradiating up to 7 kGy, and consider-ably lowered after irradiating at 7 kGy. The concentration of 50% inhibition of ovalbumin to IgG was increased to 1.5~3.7 times in an irradiation dose-dependent relationship. SDS-PAGE of ovalbumin showed that the partial breakdown of ovalbumin was induced by irradiation. The lowering of binding activity was probably due to the partial breakdown of ovalbumin by irradiation. These results demonstrated that the ELISA should be quite useful and effective methods for the identification of irradiated eggs.

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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.

학술자료I- 고양이 외인성 췌장부전 Part 2(Feline Exocrine Pancreatic Insufficiency)

  • Lee, Jin-Su
    • Journal of the korean veterinary medical association
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    • v.47 no.12
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    • pp.1086-1090
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    • 2011
  • 외인성 췌장부전은 췌장에서 분비되는 소화효소의 부족한 합성 및 분비에 기인한다. 대부분의 개체에서 보이는 임상증상은 체중감소, 다량의 연변, 지방변이다. 혈청 고양이 유사트립신 면역활성 농도검사는 외인성 췌장부전 진단에 있어 매우 유용한 검사법이다. 외인성 췌장부전 환자의 치료는 췌장 소화효소의 공급이다. 대부분의 외인성 췌장부전 환자에서 심각한 혈청 코발라민의 감소와 동반되기 때문에 코발라민의 투여는 반드시 필요하다. 마지막으로 개에 비해 흔하게 확인되진 않지만, 만성적인 연변 및 체중감소를 보이는 환자에서는 진단에 있어 외인성 췌장부전에 대한 고려가 필요하다.

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PNMT유전자 조각의 shot-gun cloning에 관한 연구

  • 조홍범;최영길
    • The Microorganisms and Industry
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    • v.16 no.1
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    • pp.21-26
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    • 1990
  • 동물체의 신경전달물질은 acetylcholine을 비롯한 9종 이상의 물질이 알려져 있으며 이들 물질의 생체내에서의 기작과 생합성 과정에 관해서는 많이 알려져 있고 또한 이 방면의 연구도 활발하다. 그중에도 Joh와 Baetge등에 의하여 DBH와 PNMT 효소의 생산에 관여하는 mRNA가 면역침전법에 의하여 순수 분리되고 이를 주형으로 하여 DBH와 PNMT 효소 생산에 근본이 되는 유전자의 염기서열 및 유전자의 구조를 규명하는 작업이 진행되고 있으나 아직도 그 전체가 규명된 바는 없다 (Baetge등, 1981;1983;Joh등, 1983;1984). 그리하여 본인들은 상기의 연구자들로부터 PNMT 유전자인 cDNA를 M13mp18과 19의 vector phage에 재조합시키고 이 cDNA를 JM107rhk 109의 host bacteria에 도입하여 형질발현 실험을 통하여 확인하고자 하였다.

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Detection of bovine rotavirus antigen by enzyme linked immunosorbent assay (효소면역법에 의한 소 로타바이러스 항원 검출)

  • 안재문;유기조;이용희;이종인
    • Korean Journal of Veterinary Service
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    • v.19 no.1
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    • pp.30-38
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    • 1996
  • Enzyme-linked immunosorbent assay (ELISA) was developed to detect rotavirus antigen in fecal samples using VP6-specific monoclonal antibody(2B12). The ELISA for rotavirus antigen detection found to have specificity to all bovine and porcine rotaviruses tested but not to bovine viral diarrhea virus and bovine coronavirus. The ELISA appeared to have similar sensitivity and specificity compared to fluorescence antibody assay(FA) and electropherotyping (PAGE).

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간장내 Superoxide Dismutase 측정법

  • 임동윤;고석태
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.149-149
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    • 1993
  • 목적:동식물세포에는 superoxide(0$_{-2}$)의 불균화 반응을 촉매하는 superoxide dismutase (SOD)가 존재한다. 이 효소의 생리적 의의는 지금까지도 명확하게 되어있지 않는면이 많지만, 그 의의를 명확하게 하기위해서도 측정법의 확립이 필요하다. 방법: SOD측정 방법으로는 1) Cytochrome C method 2) Nitroblue tetrazoliun method (NBT법) 3) 면역학적 방법 4) 화학발광법 등이 있다. 실험 재료는 흰쥐, mouse, 토끼의 간을 이용하였으며, 또한, 노화 및 암세포를 이용한 방법을 이용하였다. 결과: Cytochrome C 방법을 통해서 각 장기조직 (신장, 간장, 폐)에서 SOD를 측정하였으며 SOD 활성이 낮은 암조직이나 배양세포에서는 NBT 방법이 측정방법으로 적합한 것으로 나타났으며 ,간장세포내에서의 SOD의 존재부위를 확인하는 방법으로는 면역 황금 표지방법을 사용하므로 간장 mitochondria 내에 Cu, Zn-SOD가 존재함을 알 수 있었다.

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Property Comparison of Polyclonal Anti-Soy Protein Antibodies Produced for ELISA (효소면역측정법을 위한 다클론 항대두단백 항체의 생산 및 특성비교)

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Yun, Sung-Seob
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1221-1226
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    • 2000
  • Specific antibodies were produced to develope the enzyme-linked immunosorbent assay for analysis of soy proteins and the properties of the antibodies were compared. Isolate soy protein(ISP), and ISP heated with SDS and urea (ISP(SU)), acidic subunits(AS) of 11S globulin were immunized to produce polyclonal antibodies. By using competitive indirect ELISA(ciELISA), the reactivities of the antibodies toward soy proteins treated with different methods were investigated and shown as $IC_{50}$. $IC_{50}'s$ of anti-ISP antibodies to ISP, ISP(SU), ISP treated with 2-ME(ISP(ME)), and crude 11S were 20, 30, 36, and $1000\;{\mu}g/mL$, respectively. And the values of anti-ISP(SU) antibodies to the same antigens were 100, 5, 4, and $220\;{\mu}g/mL$ and those of anti-AS antibodies were 20, 2, 2.5, and $200\;{\mu}g/mL$, respectively. Therefore, anti-AS antibodies showed the highest reactivities toward soy proteins among the produced antibodies as determined by ciELISA.

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