• Title/Summary/Keyword: Immunodiffusion test

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Studies on serological tests for pullorum disease (추백리의 혈청학적 진단법에 관한 연구)

  • 김정태;심항섭;김태종;고태오;우종태;유기승;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.3
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    • pp.313-323
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    • 1998
  • In order to establish a sensitive and specific diagnostic method for detection of antibody to Salmonella pullorum, a enzyme-linked immunosorbent assay(ELISA) was designed and standardized. The diagnostic efficacy of the established ELISA was compared with that of the serum plate agglutination test and immunodiffusion test for pullorum disease. 1. The chicken hyperimmune sera to Salmonella pullorum, S gallinarum, S typhimurium and S typhi were shown the cross reaction to S pullorum antigen by serum plate agglutination test. 2. When compared the cross reaction titer of microplate agglutination test for chickens hyperimmune sera, it was found that the titer were 64 in S pullorum, 32 in S gallinarum, 4 in S typhimurium and 8 in S typhi, respectively. 3. When compared the specificity of various antigen(HA, EA, PA and SA) by the immunodiffusion test, the most suitable antigen was phenol-treated bactrium. 4. The optimal concentration of S pullorum antigen for ELISA was 1 : 160 dilution of bacterium. 5. The efficacy of the ELISA for detection of S pullorum antibody was compared with serum Plate agglutination test and immunodiffusion test in chickens infected with S pullorum. The antibody was first detected at 6 days after infection using three tests examined. The antibody was alldetected at 9 days by ELISA, at 12 days by serumplate agglutination test, at 15 days by immunodiffusion test.

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Survey on Actually Infected Condition of Aujeszky′s Disease to the Consigned Pigs in Seoul from 1990 to 1993 (90~93년도 서울 지역에 출하된 돼지의 Aujeszky′s병 감염 실태 조사)

  • 최준식;육동현;김성삼;문현칠
    • Korean Journal of Veterinary Service
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    • v.17 no.1
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    • pp.19-24
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    • 1994
  • The porcine Aujeszky's disease was surveyed by Enzyme Immunodiffusion method and serologic neutralization test to the slaughtered pigs at slaughtehouse only in Seoul from March, 1990 to October, 1993. After detecting the positive by enzyme immunodiffusion method primary, we decided finally the positive by serologic neutralization test secondary. Results obtained through the experiments were summarized as followed; 1. The positive of Aujeszky's disease in March, 1990 was 2 of 1,000 sera. 2 positive were decided as the consigned pigs in Kalsan, Hongsung, Chungnaa 2. The positive of Aujezsky's disease in May, 1991 was decided 1 serum at Pogok, Yongin, Kyeonggi. 3. All of the positive detected by Enzyme immunodiffusion Method in 1993 were decided finally in the negative sera. 4. The positive sera detected in 1992 were decided 32 sera at Gyeonggi, 6 at Chungnam, and 1 at Gangwon. Especially, the positive sera percentage detected by Kit Latex Aujeszky Test appeared 78.04% at Gyonggi and by enzyme immunodiffusion Method appeared 11.11% at Chungnam and Gangwon.

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Detection of antibodies in swine serum to Aujeszky's disease virus using agar-gel immunodiffusion test (Agar-gel immunodiffusion test를 이용한 돼지 혈청중 Aujeszky's disease virus 항체 검출에 관한 연구)

  • Cho, Hyo-gueon;Jun, Moo-hyung
    • Korean Journal of Veterinary Research
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    • v.30 no.3
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    • pp.297-307
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    • 1990
  • To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.

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Changes of haemolymph proteins in Pieris rapae L. during the cuticle formation and hardening process (배추흰나비의 큐티클 形成과 硬化에 따른 혈림프 단백질의 變化)

  • Hak Ryul Kim;Eul Won Seo
    • The Korean Journal of Zoology
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    • v.23 no.1
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    • pp.1-12
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    • 1980
  • Changes and possible origin of haemolymph proteins during the cuticle formation and hardening are determined by means of acrylamide gel electrophoresis and immunodiffusion. The results by acrylamide gel electrophoresis showed at least 19 protein bands in the haemolymph and 13 fractions in the fat body with relatively constant pattern during the period of cuticle formation and hardening. Both haemolymph and fat body proteins are generally characterized by the presence of three to four heavy stained bands and several thin bands near the top region of the gel. At least over five haemolymph proteins are constantly present during this period. Immunodiffusion tests show that of total eight to nine pupal haemolymph proteins two proteins were already detected in the fat body before pupation and other two proteins were also found in the fat body immediately after pupation, suggesting fat body as possible source of these two haemolymph proteins.

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Induction of Cd-binding High Molecular Weight Protein in Rat Tissues (흰쥐 조직에서의 카드뮴 결합 고분자량 단백질의 유도)

  • Chun, Ki-Jung;Kim, Bong-Hee;,
    • YAKHAK HOEJI
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    • v.41 no.3
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    • pp.352-358
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    • 1997
  • The study was carried out on the biochemical characters of Cd-BP(I) after isolation and purification of the protein from the liver of rat injected intraperitoneally with Cd. A c ontinued study has been doing whether Cd-BP(I) could be induced by Cd or by other metals such as Zn and Cu. Antisera were made against the Cd-BP(I) from NewZealand white rabbits. Carried out were ${\gamma}$-globulin purification, then Ouchterlony test adn gel immunodiffusion test. Cd-BP(I) was also found in normal tissues of rat. It was induced up to a considerable level by Cd, whose induced level was higher than that of Cu or Zn treatment. The level of induction by Cu or Zn pretreatment plus Cd treatment was lower than that by simple treatment of Cu or Zn. Such a result was presumably related to the Cd toxicity.

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Induction of Cd-binding High Molecular Weight Protein [Cd-BP(II)] in Rat Tissues (흰쥐 조직에서의 카드뮴 결합 고분자량 단백질 [Cd-BP(II)]의 유도)

  • 천기정;김봉희
    • YAKHAK HOEJI
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    • v.43 no.5
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    • pp.591-597
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    • 1999
  • The study was carried out on the biochemical characters of Cd-BP(II) after isolation and purification of the protein from the liver of rat ip injection with Cd. A continued study has been doing whether Cd-BP(II) could be induced by Cd or by the other metals such as Zn and Cu. Antisera were made against the antigen of Cd-BP(II) from New Zealand white rabbits. We carried out g-globulin purification, then Ouchterlony test and gel immunodiffusion test. Cd-BP(II) was also found in normal tissues of rat. It was induced up to a considerable level by Cd, whose induced level was higher than that Cu or Zn treatment. The level of induction by Cu or Zn pretreatment plus Cd treatment was lower than that by single treatment of Cu or Zn. Such a result was presumably related to the Cd toxicity.

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Seroepizootiological survey on bovine leukosis of dairy cattle in Kyunggi province (경기도 지역 유우의 소백혈병 항체 분포 조사)

  • 심항섭;국정희;황영옥;정봉수;김학열;이모란;유성종;강순근;임경애
    • Korean Journal of Veterinary Service
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    • v.21 no.3
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    • pp.255-260
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    • 1998
  • Since bovine leukosis caused considerable economic loss to the dairy industry, seroepidemiologi-cal survey on bovine leukosis was carried out for the dairy herds in Kyunggi province. 1. When compared the results of immunodifussion test with those of enzyme-linked immunosorbent assay(ELISA) for 94 dairy herds sera, the relationship between the immunodifussion test and ELISA were showen high corresponding rate with sensitivity(97.5%) and specificity(92.6%). 2. In immunodiffusion test for bovine leukosis virus (BLV) antibody in 570 dairy cattle from 30 herds, mean positive rate for BLV antibody was 28.2%. The positive rate by districts were 16.5% in central, 35.4% in east, 17.3% in west, 29.1% in south, 31.6% in north, 43.7% in northeast. 3. When the results of serological studies was analyzed by age groups, the number of positive was increased gradually with the advanced in age of herds. The highest positive rate was found in the age over 6 years. 4. Of 30 dairy herds examined, 5 herds(16.7%) have no reactions against BLV antigen while 15 herds (50%) showed the range of 1∼5 positive cattle and 5 herds(16.7%), the rang of over 11 positive cattle.

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A Study on the Antigen Characteristics of Rhodotorula rubra (Rhodotorula rubra의 항원특성에 관한 연구)

  • Kwon, Hyuk-Ku;Lee, Jang-Hoon;Ryeon, Kon
    • Journal of Environmental Health Sciences
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    • v.28 no.5
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    • pp.28-34
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    • 2002
  • Antigenicity of Rhodotrula rubra isolated from pulmonary tissue of pulmonary tuberculosis patients was studied by means of agglutination reaction with R. rubra whole cell antiserum. And the serological reactivity of crude polyfac charide from R. frubra, Candida albicans, Candida tropicalis, Candida, glabrata, and Saccharomyces cerevisiae ATCC 26603 with antiserum to R. rubra whole cell was studied by means of immunodiffusion test. R. rubra showed stationary phase after 48h when it was cultured in GYEP broth. While agglutinogen titer was 1:64 at lag phase, agglutinogen titer was 1 :256 after 20h. After growth of R. rubra on different 11 media, nutritional environment showed similar agglu-tination reartivity. The agglutinogen titer of C. albicans, C. tropicalis, C. giabrata, which were isolated from patient's expectoration, to R. rubra antiserum by means of agglutination reaction were 1:16, respectively. But, Sacch. cervisiae ATCC26603 was negative. Those results were lower than that of R. rubra agglutinogen titer 1:256. As a result of immu-nodiffusion test with crude polysaccharide extracted from cell wall of R. rubra, C. albicans, C. tropicalis, C. glabrata, Sacch. cervisiae ATCC26603, precipitin line was found only with R. rubra, of which antibody titer was 8.

Studies on enzootic bovine leukosis II. Survey for antibodies to bovne leukemia virus in the Holstein calves in a dairy farm (축우의 유행형 (지방병성) 백혈병에 관한 연구 II. 한 유우군에서 출생한 송아지에 대한 우백혈병 바이러스 항체 검사)

  • Kim, Chan-ju;Son, Jae-young;Ko, Ki-whan
    • Korean Journal of Veterinary Research
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    • v.30 no.3
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    • pp.343-348
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    • 1990
  • Total 51 calves born from both 28 seropositive and 23 seronegative dams were subjected to study both prenatal and postnatal infections of bovine leukemia virus (BLV), and the duration of passive colostral antibody by means of immunodiffusion (ID) test. All calves were tested for precolostral and postcolostral periods by 16 months of age. The results were as follows: 1. Of 28 precolostral sera of the calves born from infected dams, one appeared positive, indicating in utero BLV infection from the dam. 2. BLV-antibody test for the postcolostral sera of the calves born from seropositive or seronegative dams showed that the colostral antibody of the calves disappeared from 2 to 6 months of age, and the increase of the number of seropositive calves initiated from 3 to 4 months of age indicated postnatal infection.

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Indirect ELISA Method for Measurement of Lactoperoxidase using IgY Antibody (IgY 항체를 이용하여 Lactoperoxidase 정량을 측정하기 위한 Indirect ELISA 방법의 개발)

  • 이승배;최석호;최재원
    • Food Science of Animal Resources
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    • v.24 no.2
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    • pp.182-188
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    • 2004
  • To determine the concentration of Lactoperoxidase (LPO), an indirect enzyme-linked immunosorbant assay(ELISA) was developed. Anti-LPO egg yolk immunoglobulin(IgY) was transferred to egg yolk by immunizing of Brown hens with LPO. The titer of purified anti-LPO IgY was 1: 520,000. The immunological response of anti- LPO IgY with ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein and lysozyme were evaluated, resulting that the anti-LPO IgY found to be a specific antibody toward LPO and no cross-reaction was observed against ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein, and lysozyme in double immunodiffusion test and ELISA test. In indirect ELISA method, coating concentration of LPO and dilution rate of anti-LPO IgY was 0.25$\mu\textrm{g}$/mL and 1:8,000 respectively. Sensitivity in the standard curve of LPO was ranged from 0.01 to 1$\mu\textrm{g}$/mL using anti-LPO IgY.