• Title/Summary/Keyword: Immunodiffusion test

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Simultaneous Purification of Enterotoxin A and C by Fast Protein Liquid Chromatography (FPLC에 의한 Staphylococcal Enterotoxin A와 C의 동시분리)

  • Lee, Jung-Hee;Kim, Jong-Bae;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.856-861
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    • 1988
  • A new method developed for simultaneous purification of enterotoxin A and C from Staphylococcus aureus strain L 350/1 consisted of chromatography on carboxymethyl (CM)-cellulose using a buffer of variable pH, gel filtration on Ultro gel, and fast protein liquid chromatography(FPLC) using a buffer of variable pH. The enterotoxin A and C were purified by three steps: batchwise adsorption from culture supernatant on Amberlite CG-50; chromatography on CM-cellulose using a buffer of constant pH and molarity; and gel filtration on Sephadex G-75. The purified enterotoxin appeared homogeneous by gel diffusion and polyacrylamide gel electrophoresis. Upon treatment with CM-cellulose using a elution of variable pH, enterotoxin A and C were so close that they were not separated completely. After elution from gels, the enterotoxins appeared as a single peak at the same position. Gel filtration gave a reaction of complete identity to enterotoxin A and C in Ouchterlony immunodiffusion. In FPLC using a CM-cellulose, enterotoxin A and C were simultaneously separated at pH 8.6 and 6.8. When each fraction was performed to gel immunodiffusion, at peak of enterotoxin A and C were not detected each other. In a method of elution by pH-gradient was to be more efficient as a simultaneous separation method in terms of speed, yields and simplicity. The purified toxin A and C were identical to type A and C reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion.

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A Study on Antigenicity and Immunodepressive Activity of DA-125, A New Anthracycline Anticancer Agent (새로운 Anthracycline 항암제 DA-125의 항원성 및 면역독성에 대한 연구)

  • 백남기;강경구;김옥진;안병옥;이순복;김원배;양중익;정세영
    • Biomolecules & Therapeutics
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    • v.1 no.2
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    • pp.236-243
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    • 1993
  • Immunologic potential of DA-125, a new anthracycline antitumor antibiotic, was investigated using guinea pigs and mice. In antigenicity experiments, guinea pigs were sensitized subcutaneously with DA-125 or DA-125 incorporated in complete Freund's adjuvant (CFA) once a week for three weeks. No systemic anaphylaxis was induced by intravenous injection of DA-125 or DA-125 incubated with guinea pig serum after 3 weeks from the last sensitization. None of sera of these animals showed any passive cutaneous anaphylactic reaction (PCA) when DA-125 or DA-125 incubated with guinea pig serum was used as a challenging antigen in homologous PCA experiment. On the other hand the treatment of guinea pigs with ovalbumin Incorporated in CFA induced systemic anaphylactic reaction when challenged by intravenous injection of 5 mg/body of ovalbumin. Immunodiffusion test revealed no precipitating antibodies as detected in guinea pigs sensitized with DA-125. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with DA-125 or DA-125 mixed with aluminum hydroxide gel (Alum), None of sera showed positive reaction when DA-125 or DA-125 incubated with rat serum was used as a challenging antigen. Sera of animals immunized with a mixture of ovalbumin and alum showed positive PCA reaction when 5 mg/body of ovalbumin was injected as a challenging antigen. In lymphocyte proliferation tests, spleen lymphocyte proliferation to PHA and LPS was similarly impaired by 12 mg/kg of DXR or 36 mg/kg of DA-125, and the immunodepressive activity of DA-125 showed a dose-dependent manner. From these results, it could be concluded that immunosupression of DA-125 would be comparable to that of DXR and that DA-125 would not induce systemic allergic reaction in its clinical use.

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Survey for Antibodies to Bovine Leukemia Virus in Dairy Cattle (유우(乳牛) 백혈병(白血病)바이러스에 대(對)한 혈청항체(血淸抗體) 조사(調査))

  • Park, No-chan;Choi, Won-pil
    • Korean Journal of Veterinary Research
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    • v.26 no.1
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    • pp.61-68
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    • 1986
  • This paper described the distribution and transmissibility of BLV(bovine leukemia virus), the relationship between antibodies against BLV and lymphocyte count in 313 dairy cattle from 36 herds, the clinical signs and hematological findings of 2 lymphosarcomatous cattle in the northern area of Kyungpook. Eighty three (26.5%) of 313 cattle from 36 herds were positive for BLV antibodies and 19 (52.8%) of 36 herds were infected with BLV by the immunodiffusion test with BLV-gp antigen. The rate of BLV infection in cattle varied from 9.5 to 87.5% in 19 positive herds, it was higher in herds pastured during summer and included lymphosarcomatous onset than the other and also higher with the age. Eight (88.9%) out of 9 cattle which showed persistent lymphocytosis by the hematological test were positive for BLV antibodies. After 5 to 14 months, 13 (31.0%) of 42 cattle being negative for BLV antibodies in the positive herds converted into positive. Two lymphosarcomatous cattle were identified to be EBL (enzootic bovine leukemia) by the clinical sign, hematological examination and serological test.

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The changes of the plasma protein and the complements ($C_{3}$, $C_{4}$) after open heart surgery (개심수술후 혈장 단백 및 보체 ($C_{3}$, $C_{4}$)의 변화상 추적)

  • 남충희
    • Journal of Chest Surgery
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    • v.19 no.4
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    • pp.558-562
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    • 1986
  • The extracorporeal circulation has been much improved recently, but has yet much complex problems such as the protein denaturation and the activation of the complement system by the exposure of the blood to the foreign surface, which may result in such as the postperfusion syndrome. We studied the changes of the plasma protein fractions by the electrophoresis and the complement consumption [C3, C4] by the immunodiffusion method in the patients undergoing cardiac operation from Mar. 1, 1986 to Aug. 31, 1986. The results were summarized as follows: 1. y-globulin fraction was decreased [p<0.02 by paired t-test, N=25], but a,-globulin was increased [p<0.001 by paired t test, N=25] after operation. 2. C3,C4 were significantly reduced [p<0.001 by paired t-test, N=14] postoperatively and normalized from 24 hours after operation. 3. The consumption of C3,C4 had significant linear correlation [correlation coefficient r=0.97] and C, was more markedly reduced comparing with C3, which probably means the complement activation by classical pathway in our bubble oxygenator group.

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

  • Kim, Jong-man;Ahn, Jong-sam;Woo, Seung-roung;Jo, Dong-hee;Jo, Yun-sang;Park, Jeung-moon;Yoon, Yong-dhuk;Chang, Guk-hyun
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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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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ACUTE INFECTIOUS BURSAL DISEASE IN CHICKENS : PATHOLOGICAL OBSERVATION AND VIRUS ISOLATION

  • Chowdhury, E.H.;Islam, M.R.;Das, P.M.;Dewan, M.L.;Khan, M.S.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.4
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    • pp.465-469
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    • 1996
  • Pathological and virological investigations were conducted on suspected outbreaks of infectious bursal disease (IBD) in a broiler farm and five pullet-raising poultry farms of Mymensingh and Tangail districts of Bangladesh. About 80 to 100 percent chicks were affected at the age of 26 to 45 days and mortality varied from 20 to 30 percent in broilers and 40 to 80 percent in layer chicks. Signs, symptoms, gross and microscopic lesions were typical of acute IBD. Several isolates of virus could be obtained by embryo inoculation and the virus was diagnosed as infectious bursal disease virus (IBDV) by agar gel immunodiffusion test (AGID). The virus isolate belonged to the very virulent pathotype of IBDV causing 100 percent mortality in three weeks old chicks on experimental infection.

Characterization of Mosquitocidal Bacillus thuringiensis Strain H9B (모기유층에 대한 살충성 Bacillus thuringiensis H9B 균주의 특성)

  • 이기희;김광현;김병우
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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    • pp.393-398
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    • 1993
  • One strain of mosquitocidal Bacillus thuringiensis, H9B, was isolated from soil. The biochemical characteristics and flagella antigenicity of the strain H9B is similar to that of B. thuringiensis subsp. darmstadiensis. The delta-endotoxin of the strain H9B coincided with that of B. thuringiensis subsp. darmstadiensis strain 73E10-2 on agarose double immunodiffusion test. The delta-endotoxin of B. thuringiensis subsp. israelensis contains hemolysin fragment (28 kb) on SDS-PAGE when the delta-endotoxin was solubilized in alkali, while that of the strain H9B does not contain 28 kb protein.

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Pathological and serological detection of bovine viral leukosis in a dairy farm in Jeonbuk province (유우농장에서 발생한 소바이러스성 백혈병의 병리학적 및 혈청학적 조사)

  • Jo Young-Suk;Jang Sae-Gun;Chu Keum-Suk;Choi Eun-Young;Chon Hee-Woong;Hong Jae-Hee;Lim Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.29 no.2
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    • pp.89-96
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    • 2006
  • Bovine viral leukosis is a viral disease of cattle characterized by the development of tumors in the lymphatic tissue. A female Holstein, 3-year-old, was submitted for diagnosis at the Diagnostic laboratory, Chonbuk National University. Clinical sign of the affected animal showed emaciation, enlargement of superficial lymph node and mild diarrhea. Remarkable lesions were enlargement of many internal lymph nodes. Histopathology revealed excessive neoplastic lymphoid cells characteristic of BVL infection. Subsequently, serums from all cattle were collected and serological examination was done where a 85% seropositive rate was detected using ELISA test. ELISA method showed a comparatively 75% higher detection rate than the agar gel immunodiffusion (AGID) test (85% vs 40%). Serologically positive cattle were variably detected in all ages from under 1 year to over 6 year of age. Hematological examination consistently showed leukocytosis and a differential lymphocytosis of seropositive cattle. Detailed comparative pathological and serological data diagnosed the presence of bovine viral leukosis.

Enzymeimmunoassay for the Plasma Vitellogenin and Early Determination of Ovarian Maturation in Red Seabream, Pagrus major (참돔(Pagrus major)의 혈장 난황단백전구체에 대한 효소면역측정법과 난소성숙의 조기판정)

  • Han Chang-Haa;Yang Mun-Ho;Paek Jae-Min;Lim Sang-Koo;Kim Kwang-Hyun
    • Journal of Aquaculture
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    • v.8 no.1
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    • pp.1-19
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    • 1995
  • In red seabream, Pagrus major the female specific protein in the vitellogenic female serum was identified by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The female specific serum protein might be vitellogenin based on the results of the immunological analysis for the male and vitellogenic female sera and crude egg extracts. Also, it was identified by the immunodiffusion test that the purified yolk protein from ovarian egg extracts has antigenic identities shared with the female specific serum protein. To study the relationship between the maturational stages of gonad and plasma levels of vitellogenin, these were measured from the late resting period (January) to the vitellogenic preiod (April) by the modified enzymeimmunoassay (EIA) using antiserum against yolk protein. The level of plasma vitellogenin began to increase in February (previtellogenesis stage) and continuously increased with the ovarian growth during the vitellogenesis period (March to April). The plasma vitellogenin levels were significantly different between the females and the males in February. Validation for the modified EIA system. was tested .The absorbance curve of serial dilutions of serum from the vitellogenic female was paralleled to the standard curve of yolk protein; $109\pm5.6\%$ recovery was achieved by the modified EIA. And the intraassay coefficients of variation were less than 10% within the concentration ranging from 31.3 ng/ml to 1,000 ng/ml. These findings suggest that the sex determination in adult red seabreams could be possible by using the modified EIA as early as in February.

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Application of monoclonal antibody to develop diagnostic techniques for infectious bovine rhinotracheitis virus. II. Diagnosis of infectious bovine rhinotracheitis by using monoclonal antibody (소 전염성비기관염(傳染性鼻氣管炎) 바이러스에 대한 monoclonal antibody 생산(生産)과 진단법(診斷法) 개발 II. Monoclonal antibody를 이용한 소 전염성비기관염(傳染性鼻氣管炎)의 진단(診斷))

  • Jun, Moo-hyung;Kim, Duck-hwan;An, Soo-hwan;Lee, Jung-bok;Min, Won-gi
    • Korean Journal of Veterinary Research
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    • v.29 no.1
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    • pp.27-35
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    • 1989
  • To develop more specific and sensitive diagnostic methods for infectious bovine rhinotracheitis, 7-C-2 monoclonal antibody specific to polypeptides of infectious bovine rhinotracheitis virus (IBRV) was applied in indirect immunofluorescence antibody assay (IFA), indirect immunoperoxidase assay(IPA) and radial immunodiffusion enzyme assay (RIDEA). It was found that IBRV infected in MDBK cells could be detected as early as 8 hours post infection by IFA, and that IFA was more rapid and specific to identify IBRV antigen than IPA. The diagnostic efficacy of RIDEA and SN test was studied with 88 bovine sera. It was evident that RIDEA could eliminate the false positive reaction encountered in serum neutralization(SN) test, being more rapid and sensitive than the latter. Highly significant correlation coefficiency (r=0.76, p<0.01) was evaluated between the titers of sera and the diameters of RIDEA. Tracheal membranes and sera collected from 96 slaughtered cattle with lesions in respiratory organs were examined to detect IBRV antigen and antibody by IFA, RIDEA and SN test. It was presented that positive rates were 32.3% in IFA, 20.8% in RIDEA and 21.9% in SN test, and that coincidence rate between RIDEA and SN test were 100% in positive sera and 98.7% in negative sera. In conclusion, it was assumed that application of monoclonal antibody could improve the diagnostic efficacy of IBR by enhancing sensitivity and specificity of IPA, IFA and RIDEA.

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