• 제목/요약/키워드: agglutination

검색결과 317건 처리시간 0.026초

Partial Purification of Lectin from Mycoparasitic Species of Trichoderma

  • Singh, Tanuja;Saikia, Ratul;Arora, Dilip K.
    • The Plant Pathology Journal
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    • 제21권4호
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    • pp.301-309
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    • 2005
  • Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.

Latex 응집반응을 이용한 동물의 톡소플라즈마병 진단액 개발에 관한 연구 (Development of antigen for the microplate latex agglutination test on toxoplasmosis in animals)

  • 서명득;이응구
    • 대한수의학회지
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    • 제33권4호
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    • pp.623-632
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    • 1993
  • This study was conducted to develop a sensitized latex-antigen for serodiagnosis of toxoplasmosis in animals. Tachyzoites of T gondii(RH-strain) harvested from mouse peritoneal cavity were purified through the filtraton of polycarbonate membrane(pore size, $3.0{{\mu}m}$, Costar Co.) and disrupted by ultrasonicator. The tachyzoite suspension was ultracentrifuged for 30 min at $60,000{\times}g(4{^{\circ}C})$ and the supernatant was used as a water-lysate antigen. Polystyrene latex particles of $0.8{{\mu}m}$ in diameter(Sigma) were used for the preparation of sensitized latex-antigen suspension. The several parameters including the preparation conditions, incubation buffer. serum dilution buffer and stability of agglutination reactions were evaluated and the results obtained were summarized as follows : 1. The antigen consisting of a water-lysate of T gondii tachyzoites was adsorbed onto polystyrene latex particles of $0.8{{\mu}m}$ in diameter by adding a latex suspension to an equal volume of diluted antigen solution and by incubating the mixture at $37{^{\circ}C}$ under different conditions. 2. The optimum incubation buffer used for the antigen sensitization was 0.1M Tris-HCl buffer(pH 8.0). 3. The optimum serum dilution buffer used for the latex agglutination test was 0.1M Tris-HCl-NaCl buffer(pH 7.4) containing 300 mM NaCl. But 0.1M Tris-HCl-NaCl buffer(pH 7.4) containing 300-600 mM NaCl, 0.5% BSA and 0.01% Tween-20 improved the agglutination pattems and cleared the background of microplate well without the effects on L.A titer. 4. The time required for antigen sensitization was 40 and 60 min in incubation buffer(pH 8.0) at $37{^{\circ}C}$. But the optimun time for antigen sensitization was min at $37{^{\circ}C}$. 5. The optimun quantity of antigen absorbed on latex particles for proper agglutination was the range of 20 to $32{\mu}g$ of latex particles. 6. The optimun concentration of the latex-antigen suspension for the proper agglutination reaction was determined as 0.2%(w/v). 7. The specificity, rapidity and simplicity of the latex-particle agglutination test suggested that it might be adaptable to large scale serum screening.

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넙치의 면역반응에 수온이 미치는 영향 (Effect of water temperature on the immune response of flounder, Paralichthys olivaceus)

  • 방종득;박수일
    • 한국어병학회지
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    • 제7권2호
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    • pp.119-126
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    • 1994
  • 넙치의 면역반응에 수온이 미치는 영향을 조사하기 위하여 넙치에 포르말린 사균 Edwardsiella tarda 로 주사면역시킨 후 수온변화에 따른 체액성면역반응을 관찰하였다. 저수온 (12, $15^{\circ}C$) 에서 항체는 면역 2~3 주 후에 나타났으며 최대 항체가는 각각 16 과 32 이었다. 그러나 고수온(20, $23^{\circ}C$) 에서는 면역 1 주후에 나타났으며 최대 항체가는 약 2,048 로 높았다. 그리고 한번 형성된 항체는 수온의 변화에 민감하게 반응하였고 자연조건에서도 유사한 경향을 보였으며 면역지속기간은 19 개월 이상으로 나타났다.

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젤라틴입자응집반응법을 이용한 국내 시판 고추종자의 Tobamovirus 검출 및 오염률 조사 (Detection of Tobamoviruses and Survey on Contamination Rate in Commercial Pepper Seeds Using Gelatin Particle Agglutination Test)

  • 한정헌;장태호;이철호;김영호;나용준
    • 식물병연구
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    • 제7권3호
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    • pp.170-174
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    • 2001
  • 시판고추 종자로부터 Tobamovirus를 검출하고 Tobamovirus 오염률을 조사하고자 gelatin particle agglutination test (GPAT)의 최적조건을 조사하였다. Gelatin 입자에 감작할 담배 모자이크 바이러스 고추 계통에 대한 ${\gamma}$-globulin G 의 최적농도는 100$\mu\textrm{g}$/$m\ell$이었고, GPAT의 바이러스 검출 감도는 효소면역항체법 및 dot immuno-binding assay와 유사하였으며, Tobamovirus 검출을 위한 종자시료의 최적 희석배수는 5배에서 25배였다. 그리고 최적화된 GPAT에 의한 국내시판 고추종자의 Tobamovirus 오염률은 평균 79.1%인 것으로 판명되었다.

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A Detection Kit for Aeromonas hydrophila Using Antibody Sensitized Latex

  • Shin, En-Joo;Lee, Soon-Deuk;Lee, Kyung-Won;Lee, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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    • pp.595-598
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    • 2000
  • Aeromonas hydrophila is a pathogen to fish as well as human. It is a food-borne disease, and causes severe mortality in fish, and sometimes severe septicemia in human. In this study, a rapid detection method using latex agglutination has been developed for A. hydrophila. Polyclonal antibodies were raised against membrane and whole cells of three isolates from rainbow trout. Among these, latex particles coated with antibodies raised against whole cells of isolate No. 2 showed the best sensitivity. With latex particles coated with this antibody, we could detect $5{\times}10^4$ CFU of A. hydrophila in 5 min. The cross-reactivity with bacteria constituting the normal intestinal microflora and other pathogens for rainbow trout was insignificant. This latex agglutination assay method produced positive reaction with all clinical isolates of A. hydrophila which were identified by species-specific PCR for 16S rRNA in A. hydrophila.

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세균성 뇌수막염에서 뇌척수액 Latex 응집 추적검사의 유용성 (Prognostic Value of Repeated Cerebrospinal Latex Agglutination Testing in Bacterial Meningitis)

  • 이수정;안소현;조수진;김혜순;이승주
    • Clinical and Experimental Pediatrics
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    • 제46권4호
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    • pp.345-350
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    • 2003
  • 목 적 : 세균성 뇌수막염은 높은 사망률과 후유증을 가져오는 심각한 소아에서의 감염성 질환 중 하나이다. 본 연구에서는 이미 알려진 뇌척수액 latex 응집검사의 진단적 가치 외에도 추적검사에서 예후 예측 인자로서 유용성을 알아보고자 하였다. 방 법 : 1997년 1월부터 2002년 6월까지 세균성 뇌수막염으로 진단받고 이화의대 목동병원에서 입원치료 받았던 25명의 환아들중 뇌척수액 세균배양검사와 latex 응집검사가 모두 양성이었던 19명을 대상으로 하였다. 의무기록에서 임상증상, 뇌척수액 소견, 방사선학적 소견 그리고 신경학적 후유증에 대해 조사하였다. 뇌척수액 추적검사는 항생제 치료 후 3, 7, 14, 21일 그리고 28일에 시행하였고, 3일째 음성이 된 환아들을 I군으로, 3일 이후에도 지속적으로 양성으로 남아있는 환아들을 II군으로 나누었다. 결 과 : 대상 환아 19명의 평균 나이는 $10.6{\pm}12.3$개월(2-33개월)이었고, 원인균으로는 H. influenzae type b 11명(57.9%), Group B Streptococcus 4명(21.1%), S. pneumoniae 3명(15.7%), 그리고 E. coli 1명(5.3%)이였다. I군은 10명(52.6%), II군은 9명(47.4%)이였다. 뇌척수액 추적검사에서 세균배양검사는 3일째 모두 음성화되었고, latex 응집 추적검사는 3일째 9명(47.4%)에서 양성이었고, 7일째 31.6%, 14일째 26.3%, 21일째 10.5%에서 양성이었고, 28일째에 모두 음성이 되었다. I군에서는 2명(20.0%)에서 경막하삼출 소견을 보였다. II군에서는 9명(100.0%) 모두에서 경막하 삼출을 보였고, 이 중에서 44.4%에서 뇌수종 또는 뇌연화 소견을 보였다. 신경학적 후유증은 II군에서 감각신경성난청 1명, 경련성 질환 1명 그리고 우측반신마비 1명이 발생했고, I군에서는 발생하지 않았다. 결 론 : 세균성 뇌수막염 치료 시 뇌척수액 latex 응집 추적검사를 실시하는 것은 치료 초기에 신경학적 예후를 예측하는데 유용하였다.

리포좀을 이용한 생리활성물질의 면역학적 분석법 (Liposome Immunoassay for Bioactive Substances)

  • 김종국;박경미
    • Journal of Pharmaceutical Investigation
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    • 제24권4호
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    • pp.201-215
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    • 1994
  • Recent development in the immunochemical technique has resulted in a new ultrasensitive analytical method known as liposome immunoassay (LIA). Liposome is a key element in performing liposome immunoassays, specifically designed to participate in immune reactions. A variety of markers can be encapsulated in liposomes and used as quantitative indicators of reactions. Liposome immunoassay based on agglutination, complement-mediated Iysis, cytolysin-mediated Iysis, detergent-mediated Iysis or destabilization of the liposomal membrane have been reviewed. The quantity of markers released from liposomes should be proportional to the concentration of the analytes. Therefore, liposomal agglutination and Iysis which are essential to liposomal Iysis are critically reviewed to provide a better understanding of liposome immunoassay. Based on the literature review of recent advances in liposome immunoassay for bioactive substances, this assay method may provide a convenient, specific and highly sensitive method for detecting and measuring trace amount of clinically relevant substances in the future.

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식물의 렉틴 성분 스크리닝 (Screening of Plants for Lectins Constituents)

  • 정시련;정수민;이승호;전경희
    • 약학회지
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    • 제40권4호
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    • pp.387-393
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    • 1996
  • The erythrocytes agglutination test was applied to the common korean plants for lectin activity screening by using human blood. During the four years, 108 species from 46 families of floras were collected, identified and subjected to the test after being divided into several different parts. Only 13 species demonstrated strong lectin activities. Meanwhile 66 species did not shown any agglutination. All others were observed as having low activity or as having hemolytic constituents.

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방사성동위원소(放射性同位元素)로 처리(處理)한 추백리균(雛白痢菌)의 응집반응(凝集反應)에 대한 비교시험(比較試驗) (A Comparative Experiments for Tube Agglutination Test of Pullorum Antiserum with gamma-Ray 60Co Irradiated Salmonella pullorum)

  • 서부갑
    • 대한수의학회지
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    • 제16권1호
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    • pp.7-10
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    • 1976
  • An agglutinability between naturally infected positive chicken serum of pullorum disease and hyperimmumized rabbit antiserum was compared. And the following results were obtained and summarized. 1. On the agglutinability, Salmonella pullorum antigen which irradiated gamma-ray was more excellent than another both formalized and heated antigen. 2. Time of judgemented as positive titer in the tube agglutination test to the naturally infected positive chicken serum was it most suitable for 12 hours at $37^{\circ}C$. 3. Agglutination titer of positive immune chicken serum against gamma-ray irradiate Salmonella pullorum were as 320~640x.

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남성불임 환자에서 Tray Agglutination Test 및 Indirect Immunobead Test를 이용한 항정자항체에 관한 연구 (Antisperm Antibodies in Infertile Males as Detected by Tray Agglutination Test and Indirect Immunobead Test)

  • 김철수;이무상;홍재엽
    • Clinical and Experimental Reproductive Medicine
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    • 제17권2호
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    • pp.153-158
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    • 1990
  • Sera from 95 infertile males males were assayed for antisperm antibodies using the Tray Agglutination Test(TAT) and indirect Immunobead Test(IBT). The correlation between antisperm antibodies and seminal analysis in infertile men was evaluated, and the TAT was compared with new indirect IBT. The results were obtained as follows. 1. Positive rate for antisperm antibodies was high in azoospermia, oligoasthenospermia, pyo-spermia, in order, Especially in obstructive azoospermia, the rate was the highest in both methods 2. Positive rates for antisperm antibodies in TAT was higher than indirect IBT. 3. Among the isotypes of the immunoglobulins, IgG were most prevalent. IgG and IgA were bound predominantly to the head and IgM predominantly to the tail up.

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