• 제목/요약/키워드: Fasciola

검색결과 95건 처리시간 0.026초

효소면역전기영동이적법을 이용한 유조설고충 단세후군항체 및 환기혈청에 대한 항원특리성 분석 (Analysis of antigen specificity using monoclonal and polyclonal antibodies to cysticercus cellulosae by enzyme-linked immunoelectrotransfer blot technique)

  • 조승열;강신영;김석일
    • Parasites, Hosts and Diseases
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    • 제25권2호
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    • pp.159-167
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    • 1987
  • 기생충질환의 진단에도 이용하는 혈청학적 진단에서는 근록기생충항원과 교우반응이 나타나 특이도를 떨어뜨리는 경우가 있어 문제가 되고 있다. 진단용으로 사용하는 기생충노항원의 단백질 구성성분중 교*반응을 가장 잘 일으키는 것이 어떤 것인지를 알 수 있다면 특이도가 높은 항원을 제작하는 기본정보로 사용할 수 있을 것이다. 최근 SDS-폴리아크릴아마이드젤 전기영동(SDS-PAGE)과 효소면역전기영동이적법(EITB)을 이용하여 항원항체반응을 일으키는 단백질성분을 노항원중에서 구별할 수 있게 되었다. 이 연구에서는 유구낭미충의 낭액, 두절의 생리식염수추출액, 전낭미충추출액과 스파르가눔추출액, 포충낭액, 간질추출액, 간흡충추출액, 폐흡충추출액 등 8종의 항원에 대하여 유약낭미충증 환자 24명의 혈청과 본교실에서 제작한 단세포군항체가 어떻게 반응하는지 관찰하여 항원특이성을 분석하고자 하였다. SDS-폴리아크릴아마이드젤 전기영동은 길이 9cm, 두께 1.5mm인 10~15% linear gradient gel을 사용하였고 시료 $30{mu}l$를 sample buffier와 합께 $95^{\circ}C$에서 5분간 가열한 후 3~4시간 전기영동하였다. 분리된 단백질을 다시 100V에서 2시간 동안 Towbin buffer에서 전기영동하여 nitrocellulose 종이로 이적하였다. 그후 환자혈청(1 : 100), 단세포군항체(1 : 100)과 반응시키고 peroBidase-conjugated antihuman IgG(또는 perozidase-conjugated antimouse IgG), 기질의 순으로 반응시켜 발색하였다. 그 결과를 요약하면 다음과 같다. 1. SDS-폴리아크릴아마이드젤 전기영동후 Coomassie brilliant blue R-250으로 염색한 바 유구낭미충 낭액에는 단백질대가 최소 23개 나타나고 그중 94, 64, 48, 39, 34, 24, 15, 10, 7kDa가 주단백질이었다. 두절추출액에는 단백질대 34개가 보였고 그중 94, 64, 39, 34, 26, 24, 21, 15, 10, 7kDa가 주단백질대이었다. 전낭미충추출액도 두절추출액과 동일한 소견을 보였다. 낭액항원과 충체추출액간에는 단백질 구성성분보다는 구성비의 차이가 심하였다. 스파르가눔 추출액은 28%, 포충낭액은 15%, 간질추출액은 22%, 간흡충추출액은 6개, 폐흡충추출액은 11개의 단백질대를 보였다. 2. 환자혈청 24개를 SDS-PAGE로 분리된 단백질대와 반응시킨 결과, 낭액항원에서는 평균 7.4개와 반응하였는 바 그중 152, 94, 72, 64, 48, 24, 15, 10, 7kDa 단백질이 가장 중요하였다. 두절 추출액과는 평균 6.3개와 반응하였고 그중 94, 64, 52, 39, 34, 15, 10kDa가 중요하였다. 스파르가눔추출액과는 평균 2.7개가 반응하였고 그중 130, 64kDa가 중요하였으며 포충낭액과는 평균 1개가 반응하였고 52, 27kDa가 중요하였다. 간질, 간흡충 및 폐흡충추출액과는 반응대가 관찰되지 않았다. 3. 단세포군항체는 낭미충낭액의 15, 10, 7kDa 및 두절추출액 10kDa와 반응하였다. 이상의 결과에서 유구낭미충의 구성단백질중 15, 10, 7kDa 등 저분자량단백질이 진단특이성이 높은 분획으로 판단하였다.

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간질증의 효소면역학적 진단 (Application of Micro-BLISA in Serodiagnosis of Fascioliasis in Cattle)

  • 이재구;백병걸;이상복
    • Parasites, Hosts and Diseases
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    • 제23권1호
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    • pp.95-101
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    • 1985
  • Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciela hepatica, perokidase of conjugate anti-cattle Is G and orthophenylenediamine as a substrate by micro-method technique of Volley et at. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows. 1. In assay for optimal dilution of stock antigen, the antigen (protein contents; 0. Bmgymz) was diluted from 1150 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y: -0.181-0.00127X in infected sera, and log Y: -0.578-0. 000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. 2. In assay for optimal dilution of sera, the sera were diluted from 1125 to 1/400 with in PBSJ Tween 20 (pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y: -0.1540-0.0007238X in infected sera and log Y: -0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). 3. In the 27 cases of negative intradermal test, OD values of the ELISA are $0.447{\pm}0.144$, the 95% confidence interval (Mean+2 H SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100% in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. 4. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. 5. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was $0.846{\pm}0.224$. The 75% (44 cattle) among them had higher value with compared to the criterion, and the 60% (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. 6. Prevalence of fascioliasis was 43.4% in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

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간질증(肝蛭症)의 효소면역학적(酵素免疫學的) 진단(診斷)을 위한 항원분획(抗原分劃) (Fractionation of Antigen for ELISA of Bovine Fascioliasis)

  • 이재구;백병걸;이존화
    • Parasites, Hosts and Diseases
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    • 제24권2호
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    • pp.171-176
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    • 1986
  • In order to obtain the most specific and sensitive antigen from crude antigens of Fasciola hepatica for the immunodiagnosis of bovine fascioliasis by the enzyme linked immunosorbent assay (ELISA), phosphate buffered saline extract of F. hepatica was prepared. The crude extract was fractionated into 7 antigens using sephadex G-100 column chromatography. Seven fractionated antigens were applied to ELISA, precipitation test and intradermal test, respectively. Results obtained are as follows: 1. The specificity (95% confidence interval in negative sera of bovine fascioliasis; Mean+$2{\times}SD$ of absorbence) of the first (MW>150,000) and the second antigens(MW 120,000) were 93.7%, but those of others including crude antigen showed 100%. 2. The sensitivity(positive sera of bovine fascioliasis having higher values with compared to the criterion) of the first, the sixth (MW 16,000) and the seventh antigen (MW<5,000) were 91.6%, 87.5% and 0%, respectively, but those of others showed all 100%. 3. The absorbance by ELISA using the fifth antigen (MW 26,000) was 8. 43-folds higher in the positive sera than that in the negative sera. This could be used as one of the most specific antigens for the immunodiagnosis of bovine fascioliasis. 4. In Ouchterlony test, precipitin lines were not found in the sera naturally infected with F. hepatica, but some were found in the sera of rabbits immunized with the crude antigens. The numbers of precipitin lines in the sera of rabbits were different in the different fractionated antigens. They were 6 in the crude, 2 in the second and the third antigens, 1 in the fourth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. 5. The wheal size for bovine infected with F. hepatica was $2.46{\pm}0.15cm$ in the intradermal test antigen(saline extract of F. hepatica) supplied by the Veterinary Research Institute, Rural Development Administration, Korea. The wheal size of the first, the second and the third antigens were larger than that. of intradermal test antigen, whereas those of the fouth, the fifth, the sixth and the seventh antigens showed smaller than that of the intradermal test antigen. The results suggest that the fifth antigen may be specific antigen for the immunodiagnosis of bovine fascioliasis.

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우췌질증(牛膵蛭症)의 피내반응(皮內反應)에 관한 연구(硏究) (Studies on the Intradermal Reaction in Bovine Eurytremiasis)

  • 김화식;박무서
    • 대한수의학회지
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    • 제14권1호
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    • pp.59-71
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    • 1974
  • The studies were conducted on the allergic skin reaction of bovine eurytremiasis. In order to confirm the appearance of allergic skin reaction in this disease, a protein fraction of Eurytrema SPP(EPF) was extracted, and it's antigenicity was studied by means of intradermal reaction on the rabbits, goats and bovine. Rabbits used in this studies were divided into three groups: the first group was sensitized with EPF, the second group was sensitized with EPF, and the third group was served as control which is consisted with 5 non-sensitized and 2 saline injected rabbits. Three healthy native goats and bovine also used. Among these, 86 cases were eurytremiasis, 31 cases were fascioliasis and 71 cases non-infected. A protein fraction was also made from adult worm of Fasciola SPP(FPF) and tested the cross reaction between these two different fluke infestations. The results obtained were as follow: 1. Intradermal reaction was appeared in the infected cattle(bovine eurytremiasis) as well as in the EPF sensitized rabbits and goats. 2. In the EPF sensitized rabbits, the wheal of the intradermal reaction reacted the maximum size (increased wheal size of 5.0 to 9.0 mm) in 40 to 60 minutes and erythema in 60 minutes after intradermal injection of EPF antigen. In the EPF sensitized goats, the size of wheal reacted the maximum(increased wheal size of 5.5 to 8.0 mm) in 30 to 40 minutes, and then disappeared in 120 to 180 minutes after injection of EPF antgen. On the contrary, in the control groups of rabbits and goats, the wheal (7.0 mm) produced following antigen injection reached only to 7.5 to 8.5 mm. in 10 minutes without developing erythema, and then perished in 40 to 60 minutes after injection of EPF antigen. In the infected cattle, the wheal of the intradermal reaction reached the maximal sizes(increased wheal size of 4.0 to 11.0 mm) in 15 to 20 minutes and perished in 60 to 90 minutes after injection of EPF antigen. 3. It would be standardized that the positive reaction is above 4.0 mm, the negative below 2.0 mm and the suspective is 3.0 mm of the infected cattle. In the results of intradermal reaction, 87.2%(75 of 86 cases) was positive, 4.7%(4 of 86 cases) negative and 8.1% (7 of 86 cases) was suspective. However, in the non-infected control cattle, the wheal produced following injection disappeared in 30 to 40 minutes without increasing the wheal size in the most subjects or with incresing only to 1.0 to 2.0 mm in a few cattle. 4. In the results of intradermal reaction which were reciprocally tested in EPF sensitized rabbits by FPF antigen and in FPF sensitized. rabbits by EPF antigen, there were cross reacted. In the 31 cases if fascioliasis tested by EPF antigen, 11 cases were not specifically reacted with EPF antigen. Judging from the results of cross reaction in experimental rabbits and natural bovine fascioliasis, it would be expected when both EPF and EPF antigens are given intracutaneously on the same bovine patients of eurytremiasis or fascioliasis, the former antigen will produce the larger wheal in eurytremiasis, while the latter antigen will produce also the larger wheal in fascioliasis.

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임지의 축산적 이용에 관한 연구 제2보. 강원도의 새마을 "소" 임간공동방목사업의 문제점과 개선책 (Studies on the Utilization of Woodland for Livestock Farming II. Problem and Its Improvement Followed by the Join Cattle Grazing in king Won Do)

  • 맹원재;윤익석;유제창;정승헌
    • 한국초지조사료학회지
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    • 제3권2호
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    • pp.100-111
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    • 1983
  • 본(本) 연구(硏究)는 강원도(江原道) 새마을 '소' 임간공동방목사업(林間共同放牧事業)의 일환(一環)으로 81년도(年度)에 개설(開設)된 105개(個)의 공동방목장(共同放牧場)과 '82년도(年度)에 개설(開設)된 103개(個)의 공동방목장(共同放牧場)의 경영실태와 분석(分析)된 문제점(問題點) 그리고 개선방안(改善方案)에 관한 연구결과(硏究結果)를 요약(要約)하면 다음과 같다. 1. 공동방목(共同放牧) 사업(事業)의 효과 1) 방목기간중(放牧期間中) 1 일(日) 평균(平均) 증체량은 0.46kg으로서 농가(農家) 관행사육(慣行飼育)의 0.33kg보다 높았다. 2) '82년도(年度) 208개(個) 공동방목장(共同放牧場)의 방목기간(放牧期間)(5-10 월(月))중(中) 임간공동방목(林間共同放牧) 사업(事業)의 효과를 경제분석하면, 관행사육(慣行飼育)보다 293,075.,300원의 증체효과, 543,838,750원의 인건비(人件費) 절감효과 및 194,443,270원의 사료비(飼料費) 절감효과를 얻어 약(約) 1,031,357,320원의 소득효과를 가져왔다. 3) 208개(個) 공동방목장(共同放牧場)의 설문(設問) 조사(調査) 결과(結果), 농가(農家) 관행(慣行) 사육(飼育)보다 공동방목장(共同放牧場) 순위별(順位別) 효과에 대해서 농민들은 첫째 노동력(勞動力) 절감(節減). 둘째 사료비(飼料費) 절감(節減), 셋째 질병(疾病) 넷째 다두사육(多頭飼育) 가능(可能), 다섯째 협동심고취(協同心鼓吹), 여섯째 증체 효과, 일곱째 사양관리(飼養管理) 용역(容易), 여덟째 시설비(施設費) 절감(節減)을 들고 있다. 2. 공동방목(共同放牧) 사업(事業)의 문제점(問題點) 1) 임간공동방목(林間共同放牧) 2년차(年次)부터는 야생초(野生草)의 재생력(再生力)이 현저하게 저하(低下)되어 풀의 부족 현상이 일어난다. 2) 임간공동방목장(林間共同放牧場) 적지(適地)가 국유지(國有地)에 많으나 산림청(山林廳)의 이용(利用) 허가(許可)가 나지 않아 이용이 불가능하다. 3) 방목(放牧)으로 인(因)하여 발정(發精)한 암소를 발견하기 어려워서 수정시기(授精時期)를 놓치는 경우가 많다. 4) 각(各) 방목우(放牧牛)에 대한 방역(防疫) 및 진료(診療)의문제점이 많다. 3. 임간공동방목(林間共同放牧) 사업(事業)의 개선책(改善策) 1) 공동방목장(共同放牧場) 2년차(年次)부터는 겉뿌림초지(草地)나 제경초지(蹄耕草地)를 조성(造成)하여 충분한 조사료(粗飼料)를 확보(確保)시킬 것. 2) 정부(政府)는 강원도(江原道) 내(內) 모든 국유지(國有地)의 방목(放牧) 적지(適地)는 임간공동방목장(林間共同放牧場)으로 이용하여 우육(牛肉) 증산(增産), 독우(犢牛) 생산(生産) 지대(地帶)로 활용(活用)되도록 조치(措置)할 것. 3) 여지(與地)의 방목장(放牧場)에는 우수(優秀) 종빈우(種牝牛)를 혼목(混牧)시켜 번식성적(繁殖成績)을 올리도록 한 것. 그리고 발정(發情) 촉진(促進) 홀몬 주사(注射)로 동시(同時) 발정(發情)을 유도(誘導)해서 일괄 수정(授精)시킬 것. 4) 방목장(放牧場)에 토양병(土壤病)인 기종저의 예방(豫防) 주사(注射), 간질충에 대한 구충제의 년간(年間) 2회(回) 투여, 진드기 방제(防除)를 위하여 약욕(藥浴)을 시킬 것. 4. 임간공동방목장(林間共同放牧場) 육성(育成)을 위한 정책방향(政策方向) 1) 정부(政府)는 전국(全國)의 임야(林野)를 대상(對象)으로 임간공동방목장(林間共同放牧場) 적지(適地)를 조사(調査)할 것. 2) 정부(政府)는 임간공동방목장(林間共同放牧場) 적지(適地)로 판단되는 지역은 국공유림(國公有林)이나 법적(法的) 제한(制限) 지역(地域)도 목장(牧場) 개설(開設)이 가능하도록 조치할 것. 3) 정부(政府)는 여지(餘地)에 있는 공동방목장(共同放牧場) 적지(適地)에는 도로(道路) 개설(開設)과 전기목붕(電氣牧棚) 시설(施設)을 정부(政府) 자금(資金)으로 지원할 것. 4) 새마을 운동(運動)의 방향(方向)을 축산소득증대(畜産所得增大)에 두고 강원도(江原道)의 특성(特性)에 맞게 계속 임간공동방목(林間共同放牧) 사업(事業)이 추진(推進)될 수 있도록 정책적(政策的)인 배려가 필요하다. 5) 정부(政府)는 공동방목장(共同放牧場) 경영에 있어서 번식(繁殖) 성적(成績) 향상(向上)을 위한 인공수정상말비점(人工受精上末備点)을 보완(補完)해 줄 것. 6) 정부(政府)는 소 값의 적정(適定) 가격(價格) 수준(水準)을 유지(維持)하기 위한 가격(價格) 정책(政策)을 실시(實施)할 것. 7) 정부(政府)는 임간공동방목장(林間共同放牧場)에서 초지조성(草地造成)의 신청(申請)이 있을 때는 우선적으로 허가(許可)해 줄 것.

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