• 제목/요약/키워드: immunodiffusion

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면역학적인 방법에 의한 한우와 유우의 요네병 발생조사 (A Survey of paratuberculosis by immunological methods in dairy and Korean native cattle)

  • 김종만;안종삼;우승룡;조동희;조윤상;박정문;윤용덕;장국현
    • 대한수의학회지
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    • 제34권1호
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    • pp.93-97
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    • 1994
  • A immunological survey of paratuberculosis in dairy and Korean native cattles was conducted by enzyme linked immunosorbent assay(ELISA), complement fixation test(CFT), agar gel immunodiffusion test (AGID) and intradermal skin test(ID). Over all prevalence of pararuberculosis in cattles was 6.7%(109/1633) by ID, 7.5(205/2719) by AGID, 9.3% (245/2641) by CFT and 13.4%(363/2719) by ELISA. Prevalence in dairy cattle was higher than that of Korean native cattle. Of 70 ELISA-positive cattle, 23(28.6%) and 48(68.6%) cattles were classified as positive in the AGID and positive or suspect in CFT, respectively. Of 92 ELISA-suspect cattle, 32(34.9%) and 48(52.2%) cattles were classified as AGID-positive and CFT-positive or suspect, respectively. It was concluded that paratuberculosis is widespread in cattle of Korea.

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누에(Bombyx mori)의 卵形成過程에 따른 Vitellogenin의 分布 및 量的 變化 (distribution and Quantitative Change of Vitellogenin During Egg Formation of Bombyx mori L.)

  • Kim, Hak-Ryul;Seo, Eul-Won
    • 한국동물학회지
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    • 제26권2호
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    • pp.83-93
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    • 1983
  • 누에의 卵形成時期동안 자성 혈림프에서 DEAE-cellullose column을 使用하여 vitellogenin을 순수하게 分離해 낸 후, 電氣泳動法과 免疫學的 方法을 使用하여 각 器官에 따른 vitellogenin의 分布와 이의 量的 變化를 調査하였다. 누에의 vitellogenin은 난성숙시기에 걸쳐 혈림프, 脂肪體 및 卵巢에 모두 分布하고 있으며, 특히 포화후 7일까지만 확인되었으며 그 이후 급격한 量的 減少를 나타내었으며, 또한 血蛋白質은 卵巢蛋白質과 最小한 3개의 同質인 蛋白質을 갖고 있는 것으로 보아 卵形成에 vitellogenin 이외의 다른 血蛋白質도 관여 되는 것 같다.

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Klebsiella pneumoniae가 생산하는 Glutamine synthetase의 정제 및 특성 (Purification and Characterization of Glutamine synthetase of Klebsiella pneumoniae)

  • 차정학;이왕식;성하진
    • 한국미생물·생명공학회지
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    • 제19권3호
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    • pp.259-264
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    • 1991
  • K.pneumoniae의 nif 유전자 발현에 미치는 glnA 산물 (GS)의 영향을 살펴보기 앞서 K.pneumoniae의 GS를 분리.정제하여 그 성질을 조사하였다. K.pneumoniae의 GS는 분자량 약 600,000의 12개 동일 subunit으로 구성되어져 이었으며 최적 반응 pH 및 온도는 각기 pH 7.0, $37^{\circ}C$ 였고 pH 5-8사이에서 활성의 변화가 크지 않았으며 $57^{\circ}C$에서 10분간 열처리 할 때 50의 활성 감소가 나타났다. 또 암모니아 농도의 변화에 따라 adenylylation-deadenylylation 기작에 의한 활성조절기작이 E.coli GS와 동일 했으며 anti-E.coli GS 항체를 이용한 immunodiffusion에서도 K.pneumoniae GS가 E.coli GS와 혈청적으로 매우 유사함이 드러났다.

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파충류 $M_4$형 젖산 수소이탈효소의 면역학적 연구 (Immunological Comparison of the Reptilian $M_4$-LDH Isozyme)

  • Park, Sang-Yoon;Cho, Dong-Hyun;Kim, Sang-Yeop
    • 한국동물학회지
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    • 제19권2호
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    • pp.79-84
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    • 1976
  • 살모사 근육조직의 $M_4$형 젖산 수소이탈효소에 대한 항혈청을 토끼에서 얻어서 척추동물 15종의 젖산 수소이탈효소와 항원-항체 반응결과를 초산셀룰로즈 전기영동법에 의하여 얻었으며, 아울러 뱀목 4종에 대한 면역확산 실험도 실시하였다. 살모사, 쇠살모사, 까치살모사 및 유혈목은 면역학적으로 동일한 젖산 수소이탈효소를 가지고 있으며 쇠살모사는 살모사속의 다른 종과는 유연관계가 먼것 같았으며 젖산 수소이탈효소 단위체의 면역학적 상이성이 파충류에서도 재확인되었다.

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

  • Lee, Yoo-Sung
    • Journal of Plant Biology
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    • 제28권2호
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    • pp.95-104
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    • 1985
  • 층층나무속(Cornus) 식물 수종과 그 근연군의 계통학적 유연관계를 밝히기 위하여 종자단백질을 이중면역확산법, 사전포화처리법, 방사상면역확산법, 면역학적 전기영동법 등으로 분석함으로 혈청학적추가자료를 얻었다. Cornus florida 면역혈청을 사용한 침전반응에서 C. drummondi와 C. stolonifera는 혈청학적 유사도가 매우 높았고, C. amomum과 C. recemosa는 다음으로 높았다. Cornus와의 근연군에 해당하는 분류군 중에서 Nyssa(Nyssaceae)가 가장 가까운 근연성을 나타냈고, Corokia (Cornaceae 또는 Saxifragaceae)와 Griselinia는 매우 낮은 근연성을 보여서 Cornaceae에의 소속을 지지하고 있지 않다. 그러나 그들의 독립된 과로서 구분하기 위해서는 많은 다른 분류학적 증거가 필요할 것으로 본다. 면역학적 전기영동법과 사전포화처리법의 조합된 기술은 비교적 정교한 종자단백질 분석이 가능했다.

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우백혈병(牛白血病) Virus 항체측정(抗体測定)을 위한 효소면역법(酵素免疫法) (Microplate Enzyme-Linked Immunosorbent Assay for Bovine Virus Antibody)

  • 최원필
    • Current Research on Agriculture and Life Sciences
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    • 제1권
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    • pp.195-199
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    • 1983
  • 우백혈청(牛白血病)virus에 대한 혈청항체의 측정(測定)을 위한 간접효소면역법(間接酵素免疫法)(ELISA)의 확립(確立) 및 ELISA법(法)의 감도(感度)를 알기 위하여 한천(寒天) gel 면역확산법(免疫擴散法)(ID)과 비교검토(比較檢討)하였으며 한우(韓牛), 유우(乳牛) 및 육우(肉牛)등 264두(頭)의 혈청(血淸)을 공시(供試)하였다. 혈청(血淸)을 가(加)하지 않은 공(孔)의 흡광치(吸光値)(C)로서 혈청치(血淸値)를 제(除)한 후, 표준(標準)BLV항체흡성혈청치(N)로서 피검혈청치(被檢血淸値)(T)를 나눈치(値)(T-C/N-C)가 1.5이상(以上)인 것을 BLV항체 양성(陽性)으로 하였을 때 gp-ID의 성적(成績)과 98.5%(259/263)가 일치(一致)되었다. gp-ID 향성혈청(陽性血淸) 145예(例)중 144예(例)가 ELISA 양성(陽性)으로 99.6%, gp-ID 음성혈청(陰性血淸) 118예(例)중 115예(例)가 ELISA음성(陰性)으로 97.5%가 일치(一致)되었다. 이상에서와 같이 gp-ID용(用)의 BLV 항원(抗原)을 사용한 ELISA법(法)을 gp-ID와 동등(同等) 또는 더 감도(感度)가 우수한 BLV의 혈청항체측정법 임이 입증되었으며 실용성(實用性)이 충분(充分)하다고 사료(思料)된다.

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Dot-immunoassay를 이용한 Aujeszky's disease virus항원 및 항체 검출 (Detection of antibodies and antigens of Aujeszky's disease virus using dot-immunoassay)

  • 전무형;조용성;장경수
    • 대한수의학회지
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    • 제34권3호
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    • pp.529-536
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    • 1994
  • To establish more specific and simple diagnostic methods for detection of the antibodies and antigens of Aujeszky's disease virus(ADV), we designed indirect dot-immunoassay(IDI) and double sandwich dotimmunoassay(DSDI) using the solid phases of nitrocellolose paper and polystyrene plate. The diagnostic efficacy of these methods was investigated. As the sensitivity of IDI was tested by various virus concentration, the specimens with the virus titer above $10^{4.0}TCID_{50}/0.2ml$ showed positive reaction, but that below $10^{1.0}TCID_{50}/ml$ revealed negative. Tonsil emulsion at the virus titer of $10^{4.5}TCID_{50}/0.2ml$ showed the highest sensitivity as diluted by 1/100. In detection of ADV antigens from the various tissues of the rats and pigs infected with ADV, IDI using monoclonal antibody showed the higher specificity as compared with IDI using polyclonal antibody and virus isolation method. The efficacy of the DSDI for detection of ADV antibody was compared with other tests. The sensitivity of DSDI was higher than virus neutralization(VN) and agar gel immunodiffusion test(AGID). Meanwhile, specificity of DSDI was lower than AGID, but similar to IDEA. In comparison with VN test, DSDI showed 96.9% agreement to VN test that is the highest of three tests. In general, application of polyclonal antibody in both tests caused the higher sensitivity but the lower specificty.

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폐암 환자에서의 폐엽절제술 전후의 혈청 Alpha 1-Proteinase Inhibitor의 변화 (Pre- and Postoperative Changes of Serum Levels of Slpha 1-proteinase Inhibitor in Patients with Bronchogenic Cancer)

  • 이재성
    • Journal of Chest Surgery
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    • 제28권3호
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    • pp.221-227
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    • 1995
  • Malignancy is one of the several exogenous and endogenous factors that increase serum alpha 1-PI. In fact, serum levels of alpha 1-PI were significantly elevated in the patients with the nonresectable bronchogenic cancer. the purpose of this work was to determine if the immediate postoperative change of serum alpha 1-PI level following tumor resection relates to the patient`s postoperative course. Clinical experimental study was carried out to investigate the postoperative changes of serum alpha 1-PI level following operation for 20 cases of bronchogenic cancer and 10 cases of control, nephrectomy patients Alpha 1-PI concentrations in serum was quantitated by use of radial immunodiffusion technique.The results were as follows ; Preoperative serum level of alpha 1-PI was significantly elevated in patients with bronchogenic cancers [p < 0.001 , when compared to normal control levels. Immediate postoperative serum alpha 1-PI level was significantly increased in patients with bronchogenic cancer [p < 0.05 , but slightly decreased at control groups. The peak serum level of alpha 1-PI was the postoperative three days, and then gradually decreased at the 5, 9, 14 days, but slightly elevated comparing to preoperative alpha 1-PI levels. Serum alpha 1-PI level in patients with adenocarcinoma was elevated, when compared to squamous cell carcinoma, but not significantly. According to the stages of the bronchogenic cancer, each levels of the serum alpha 1-PI were slightly different, but the whole postoperative changes were the general similarity. There were no significant difference in changes of the serum alpha 1-PI level, according to the operative procedures. As the alpha 1-PI is acute reactant, that it was required at the reoperative state of the bronchogenic cancer and rapid response, consumption or requirement were occurred, postoperatively. Therefore, alpha 1-PI can be perioperative indicator for the evaluation of the bronchogenic cancer.

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Guinea pig 및 mouse에 있어서 인형 과립구 콜로니 자극인자 DA-3030의 항원성 (Antigenicity of DA-3030, a Recombinant Human Granulocyte-colony Stimulating Factor, in Guinea Pigs and Mice)

  • 백남기;강경구;이순복;김원배;양중익
    • Biomolecules & Therapeutics
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    • 제2권3호
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    • pp.292-297
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    • 1994
  • This study was conducted to investigate antigenic potential of DA-3030, a recombinant human granulocyte-colony stimulating factor, in guinea pigs and mice. In the active systemic anaphylaxis test, the guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone did not show any anaphylactic reaction. In the homologous passive cutaneous anaphylaxis reaction, anti-DA-3030 antibody was not detected in guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone. On the other hand, the guinea pigs sensitized with 12.5 $\mu\textrm{g}$/heed of DA-3030 incorporated in Freund's complete adjuvant(FCA) or 1 mg/head of ovalbumin incorporated in FCA showed anaphylactic reaction. Anti-DA-3030 antibody was also detected in those guinea pigs. In immunodiffusion test using the sera sensitized with DA-3030 incorporated in FCA, precipitating antibodies were detected only in the sera sensitized with DA-3030 or DA-3030 incorporated in FCA showed. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone, none of the sera showed positive reaction. But sera of the animals immunized with 12.5 $\mu\textrm{g}$/head of DA-3030 incorporated in aluminum hydroxide gel(Alum) or 5 $\mu\textrm{g}$/head of ovalbumin incorporated in alum showed positive PCA reaction. DA-3030 did not cause anaphylactic shock or passive cutaneous anaphylaxis in guinea pigs and mice when given alone although DA-3030 incorporated in FCA or Alum induced anaphylactic shock and passive cutaneous anaphylaxis. From these results, it may be concluded the DA-3030 does not induce systemic allergic reaction when administered alone in its clinical use.

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Cloning of Xanthine Oxidase Gene from Mouse Liver cDNA Library

  • 이추희;이상일;남두현;허근
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.261-261
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    • 1994
  • Bovine milk xanthine oxidase (E.C.1.1.3.22, XO) purchased from Sigma Chemical Co. had the three protein fragments below 150 kDa on 7.5% SDS-PAGE, which did not show enzyme activity. To remove these fragments, the enzyme preparation was further purified through Sephadex G-200 column chromatography. Two peaks exhibiting enzymatic activity were separated very closely to the void volume, which were revealed as two different enzyme forms, dimeric and monomeric, confirmed by activity staining on native PAGE. Anti sera-against each of the two enzyme forms were raised by subcutaneous injection at multiple sites on the back of rabbits during 4 weeks. On the immunodiffusion test, it was found that both of the antisera of the two forms could react with each other, which implied that their epitopes were identical In the Western blot analysis of mouse liver cytosol fraction, it was found that rabbit anti-XO antibody bound well with the protein band of monomeric mouse liver XO of about 150kDa. Based on this result, mouse liver cDNA 1 ibrary was screened by in situ hybridizat ion wi th rabbi t anti -XO antibody as probe. Through the immunological screening, recombinant phages giving positive signal by the production of XO were selected and further purified. To validate these clones, purified phages were lysogenized in E. coli Y1089 and their lysates were analysed for enzyme activity and immunoreactivity, It was verified that lysates of the purified recombinant phage lysogens exhibited the enzymatic activity as well as bound wi th XO antibody, when induced by IPTG. The above results assert that selected recombinant phage carries mouse liver XO gene.

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