• Title/Summary/Keyword: agglutination

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Partial Purification of Lectin from Mycoparasitic Species of Trichoderma

  • Singh, Tanuja;Saikia, Ratul;Arora, Dilip K.
    • The Plant Pathology Journal
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    • v.21 no.4
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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.

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

  • Suh, Myung-deuk;Lee, Eung-goo
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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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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Effect of water temperature on the immune response of flounder, Paralichthys olivaceus (넙치의 면역반응에 수온이 미치는 영향)

  • Bang, Jong-Deuk;Park, Soo-Il
    • Journal of fish pathology
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    • v.7 no.2
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    • pp.119-126
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    • 1994
  • To investigate effects of water temperature on immune response of flounder, Paralichthys olivaceus, against Edwardsiella tarda, fish were immunized with formalin killed E. tarda antigen, and humoral immune response of these fish were observed. At lower water temperature (12 and $15^{\circ}C$), the antibody appeared 2 to 3 weeks after injection of formalin killed E. tarda antigen and the maximum agglutination titer was 16 and 32, respectively. However at higher water temperature (20 and $23^{\circ}C$), the antibody appeared one week after injection and the maximum agglutination titer was about 2,048. Once produced agglutination titer was sensitively responsed to variation of water temperature and showed that this phenomenon had also a similar tendency under natural condition. And it showed that agglatination titer of flounder immunized with formalin killed E. tarda maintained above 19 months.

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

  • 한정헌;장태호;이철호;김영호;나용준
    • Research in Plant Disease
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    • v.7 no.3
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    • pp.170-174
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    • 2001
  • Gelatin particle agglutination test (GPAT) was optimized for detection of Tobamovirus and contamination of the virus in commercial pepper seeds was evaluated. The optimum concentration of ${\gamma}$-globulin G, specific to tobacco mosaic virus pepper strain, was 100 ug/ml. The sensitivity of GPAT for the detection of Tobamovirus in pepper seeds was as high as enzyme-linked immunosorbent and dot immunoblotting assays. Optimum dilution ranges of the seed extract for GPAT was 5-25 folds. Using the optimized GPAT with above conditions, the rate of Tobamovirus contamination in seeds was turned out to be average of 79.1%.

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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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    • v.10 no.5
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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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Prognostic Value of Repeated Cerebrospinal Latex Agglutination Testing in Bacterial Meningitis (세균성 뇌수막염에서 뇌척수액 Latex 응집 추적검사의 유용성)

  • Lee, Soo Jeong;Ahn, So Hyun;Cho, Su Jin;Kim, Hae Soon;Lee, Seung Joo
    • Clinical and Experimental Pediatrics
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    • v.46 no.4
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    • pp.345-350
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    • 2003
  • Purpose : Bacterial meningitis is a serious infection of childhood associated with a significant morbidity and mortality. Repeated cerebrospinal fluid(CSF) examination is a useful prognostic indicator and a delayed sterilization is associated with a higher incidence of neurologic abnormalities. In this study we tried to determine the prognostic value of repeated CSF latex agglutination testing. Methods : We retrospectively evaluated 19 patients admitted to Ewha Womans University Mokdong Hospital for bacterial meningitis from January 1997 to June 2002. Bacterial meningitis was confirmed by a positive CSF culture and a positive CSF latex agglutination test. Repeated CSF examinations were done at three, seven, 14, 21 and 28 days after antibiotics therapy. Neuroradiologic studies were performed. Results : The mean age was $10.6{\pm}12.3months$(range; two to 33 months). The male to female ratio was 2.8 : 1. The causative organisms were Haemophilus influenzae type b 57.9%, Group B Streptococcus 21.1%, Streptococcus pneumoniae 15.7% and Escherichia coli 5.3%. Three days after the initiation of antibiotics therapy, repeated CSF latex agglutination tests persisted as positive in nine (47.4%) out of 19 cases, but all CSF cultures became negative. In those cases with negative latex agglutination tests three days after antibiotics therapy, neuroradiologic findings were completely normal. But, in cases with positive latex agglutination tests three days after antibiotics therapy, neuroradiologic abnormalities such as cerebral infarction, encephalomalasia occurred in 44.4%. Conclusion : Repeated CSF latex agglutination testing was valuable as a prognostic factor in bacterial meningitis. Neuroradiologic abnormalities may occur in cases with delayed clearance of CSF latex agglutination tests more often than in cases with negative latex agglutination tests three days after antibiotics therapy.

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

  • Kim, Chong-Kook;Park, Kyung-Mi
    • Journal of Pharmaceutical Investigation
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    • v.24 no.4
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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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Screening of Plants for Lectins Constituents (식물의 렉틴 성분 스크리닝)

  • Jeong, Si-Ryeon;Jeong, Su-Min;Lee, Seung-Ho;Jeon, Gyeong-Hui
    • YAKHAK HOEJI
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    • v.40 no.4
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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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A Comparative Experiments for Tube Agglutination Test of Pullorum Antiserum with gamma-Ray 60Co Irradiated Salmonella pullorum (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))

  • Seo, Boo Kap
    • Korean Journal of Veterinary Research
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    • v.16 no.1
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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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Antisperm Antibodies in Infertile Males as Detected by Tray Agglutination Test and Indirect Immunobead Test (남성불임 환자에서 Tray Agglutination Test 및 Indirect Immunobead Test를 이용한 항정자항체에 관한 연구)

  • Kim, Cheol-Soo;Lee, Moo-Sang;Hong, Jae-Yup
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.2
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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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