• Title/Summary/Keyword: cross reaction

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Studies on the Classification of Aspergillus spp. by Fluorence Antibody Reaction (형광항체반응(螢光抗體反應)에 의(依)한 Aspergillus spp.의 분류(分類)에 관(關)한 연구(硏究))

  • Moon, Hi-Joo;Kim, Sung-Kon;Lee, Bae-Ham
    • The Korean Journal of Mycology
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    • v.1 no.2
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    • pp.9-14
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    • 1973
  • Author investigated fluorence antibody reaction for the antigenic relationships between Asp niger group, Asp flavus and Asp parasiticus which was indicated as follows: 1. It was concluded that there are complete differences in the antigenic properties each other because it has not cross reaction, therefore identification of strains will be simpley classified. 2. A complete cross reaction between Asp flavus and Asp parasitic us in the Asp flavus groups existed, accordingly this reaction could not identified the strain and classified between Asp. flavus and Asp. parasiticus. 3. This experiment also followed with the separated each strains from the origin (Meju, Nuruk, ATCC, NRRL), but there no differences. From the above results, this method could be classified between Asp flavus group and Asp niger group in the genus Aspergillus, but classification of Asp. flavus and Asp. parasiticus should hardely conclude with this method.

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Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Clostridium botulinum Type F Toxin (Clostridium botulinum Type F Toxin의 면역학적 효소방법에 의한 검출에 관한 연구)

  • Lee, Jeong-Kug;K. H. Yang
    • Microbiology and Biotechnology Letters
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    • v.10 no.3
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    • pp.205-209
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    • 1982
  • The enzyme-linked immunosorbent assay using the so-called "double-sandwich"technique was applied to determine Clostridium botulinum type F toxin. Polystyrene tubes were coated with horse anti-type F toxin serum and then toxin sample was added. The tubes were subsequently treated with rabbit anti-type F toxin IgG and sheep anti-rabbit serum IgG-horseradish peroxidase conjugate. By this technique, about 10 mouse intraperitoneal 50% lethal doses (ip LD/50/) of type F toxin could be detected. Low back-ground reading was achieved with the use of phosphate-buffered saline containing 0.05% Tween 20 and 1% bovine serum albumin as diluents of rabbit IgG and conjugate. Addition of EDTA in the diluents of toxin increased ELISA extinction value significantly. No cross-reaction was observed with botulinum type A and B toxin, but type E toxin gave sleight cross-reaction.

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INDIRECT IMMUNOFLUORESCENCE FOR THE IDENTIFICATION OF ACTINOMYCES SPECIES IN PATIENTS WITH PERIAPICAL LESION (치근단 병변에서 Actinomyces 종의 검정을 위한 간접 면역형광법적 연구)

  • Chang, Won-Jung;Yoon, Soo-Han;Kwon, O-Yang
    • Restorative Dentistry and Endodontics
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    • v.21 no.1
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    • pp.121-135
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    • 1996
  • Actinomyces are Gram-positive, non-acid-fast, anaerobic or microaerophilic filamentous bacteria. These organisms are frequently detected from infected root canals and periapical lesion. The purpose of this study was to use indirect immunofluorescence to determine the prescence of select Actinomyces species in a survey of teeth associated with periapical lesion, to clarify the relationship between clinical symptoms of periapical lesions and the Actinomyces species and to study on the cross reaction among Actinomyces. Actinomyces israelii serotype I (ATCC 12102), Actinomyces israelii serotype II (ATCC 29322), Actinomyces viscosus serotype II (ATCC 19246), Actinomyces naslundii serotype I (ATCC 12104) were cultured in anaerobic condition. Rabbit antisera were prepared by intravenous injection of formalized whole cells. Indirect immunofluorescence method was used to achieve the purpose. The following results were obtained. 1. There was a relationship between Actinomyces and periapical disease. 2. A. israelii serotype I, II were frequently identified with Indirect Immunofluorescence and most often assosiated with periapical disease. In culture finding, there was no significant difference between each group. 3. Indirect Immunofluoresence is both more sensitive and more rapid than culture for identification of Actinomyces species in patients with periapical lesion. 4. A. israelii serotype I, II was highly isolated in infected root canals with local swelling, A. naslundii serotype I was highly isolated in those with foul odor, and A. israelii serotype I was found in higher frequncy in those with exudate than other bacteria. 5. In the Indirect Immunofluorescence (1 : 320), A positive cross reaction was obtained between A. israelii serotype I and A. israelii serotype II, also, A. viscosus serotype II and A. naslundii serotype I. There was no cross reaction between A. israelii serotype I, II and A. viscosus serotype II, A. naslundii serotype I.

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Study on the production and management of aquatic animals : direct detection of Edwardsiella tarda using an enzyme linked immunosorbent assay (수생산물의 생산과 관리에 관한 기초연구 : ELISA법을 이용한 Edwardssiella tarda의 직접 검출)

  • Chung, Soon-Yoon;Sohn, Sang-Gyu;Chung, Joon-Ki;Huh, Min-Do;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.10 no.2
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    • pp.75-86
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    • 1997
  • Solid phase enzyme linked immunosorbent assay (ELISA) was developed to detect the whole cells of Edwardsiella tarda from infected tissues of flounder. Cross-reaction test was performed by ELISA against fish pathogens such as A. hydrophila ATCC7966. V. anguillarum HYUFP5001, Y, ruckeri 11-4, E. ictaluri and Streptococcus sp. NG8206. Rabbit anti-E, tarda Edk-2 sera highly cross-reacted with A. hydrophila ATCC7966 and V. anguillarum HUFP5001. However, the cross-reaction was removed by using the anti-serum pre-adsorbed with A, hydrophila ATCC7966 FKC. The intra-species cross-reaction among E. tarda isolates was very high. ELISA with the whole cell antigens present in tissue homogenate appeared with highly decreased sensitivity, presumably by the co-coating of lipid or proteins in tissues. Thus, it would be necessary to use the infected tissue homogenates diluted more than 100 times with PBS for diagnosis. Interestingly, compared with the using of FKC antigen, the direct detection of viable cells in tissue homogenate showed more sensitive results with detection limit of $1{\times}10^3$ cells/ml in buffer or diluted tissue homogenate. Consequently, the ELISA method developed in this study was specific, rapid and sensitive for diagnosing edwardsiellosis.

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Studies on Leptospiral Antibody in Korean Cattle and Pigs (Leptospira 속균(屬菌)에 대한 한우(韓牛)와 돈(豚)의 혈중항체조사(血中抗體調査))

  • Suh, I.S.;Ryu, E.P.
    • Korean Journal of Veterinary Research
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    • v.12 no.1
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    • pp.91-95
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    • 1972
  • Investigation of leptospiral antibody in Korean cattle and pigs was carried out from February to October, 1971. Ten different living antigens, namely L. icterohaemorrhagiae, L. canicola, L. antumnalis, L. hebdomadis, L. australis A, L. pomona, L. pyrogenes, L. grippotyphosa, L. bataviae and L. javanica, were used. A total of 590 Korean cattle and 460 pig blood samples collected from Seoul Majang-dong slaughterhouse were tested by the rapid microscopic agglutination test. Throughout the studies the following results were obtained and summarized. 1. Of 590 serum samples of Korean cattle 51 were positive(8.64%). 2. Of 460 serum samples of pigs 27 were positive (5.87%). 3. Of 51 positive cattle samples, 29(4.92%) showed antibody to a serotype of L. icterohaemorrhagiae and 18(3.0%) to L. canicola, and 4 (0.68%) to L. pomona. Eight of L. icterohaemorrhagiae positive samples showed a cross reaction to L. canicola. 4. Of 27 positive pig samples, 14(3.04%) showed antibody to L. ictereohaemorrhagiae and 7(1.52%) to L. grippotyphosa. 4(0.87%) to L. canicola, 2(0.43%) to L. pomona. Two of L. canicola positive samples showed a cross reaction to L. grippotyphosa. 5. Serum samples of seven pigs, showing antibody positive to L. grippotyphosa were first observed in Korea. 6. Infection rate of bovine and porcine leptospirosis, in Korea, appeared to be lower than that of Japan, Taiwan, Thailand and the Philippines.

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Cross-reactivity between sera from dogs experimentally infected with Dirofilaria immitis and crude extract of Toxocara canis

  • Song, Kun-Ho;Hayasaki, Mineo;Cho, Kyu-Woan;Lee, Sang-Eun;Kim, Duck-Hwan
    • The Korean Journal of Parasitology
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    • v.40 no.4
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    • pp.195-198
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    • 2002
  • This study was performed to investigate whethere there is cross-reactivity between Dirofilaria immitis and three intestinal nematodes of dogs. In ELISA, D. immitis- infected dog sera obtained at the 4th molting stage (9-11 weeks) and microfilaremic stage (25-30 weeks) were shown to be highly reactive with crude extract of T. canis. In immunoblotting, some antigenic fractions, 44, 57 88, 100 kDa of crude extract of T. canis, were found to be positive reaction with sera of dogs infected with D. immitis. However, little or no cross-reaction were observed between sera of D. immitis-infected dogs and crude extract antigen of T. vulpis or A. caninum. These result suggest that there are partial cross reaction between sera of D. immitis-infected dogs and the antigen of T. canis.

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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Extraction and verification of highly immunologenic antigen for diagnosis of bovine brucellosis (소 브루셀라병 진단용 고면역원성 항원의 추출과 검증)

  • Bae, Jae-Hyung;Jo, Sang-Rae;Jeong, Eun-Hui;Jang, Eun-Hui;Kim, Seong-Eun;Kwon, Hui-Nyeong;Park, Dong-Yeop;Lee, Kuk-Cheon
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.135-141
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    • 2010
  • Bovine brucellosis, an important zoonosis, is diagnosed with serological tests such as the RBT, TAT using inactivated whole bacterial cells or bacterial lipopolysaccharide (LPS) antigen in Korea. However, a strong cross-reaction between Brucella spp. and Yersinia enterocolitica O9 in these tests has seriously complicated the diagnosis of animal brucellosis because Brucella spp. shares common antigenic determinants with Y. enterocolitica O9 in the smooth LPS region. In this study, Brucella-field strains were isolated from Brucellapositive Hanwoo in Kimhae, Korea and outer membrane protein (omp) which has low cross-reaction with Y. enterocolitica O9 and high immunogenicity was extracted from the field strains Then we compared ELISA using the extract with RBT-TAT. Fifteen field strains were isolated from 47 supra-mammary-lymph nodes, which were collected from 18 farms. Isolation rate was 32%. Brucella-specific antigen was identified by performing SDS-PAGE or Western blotting on extracted omp with at 0.5% n-lauroylsarcosine One hundred and ninety-two serum-samples were used in the experiment: 142 negative and 50 positive samples verified by RBT-TAT. According to ELISA results, 127 samples were negative and 15 appeared positive among 142 negatives by RBT-TAT, while 42 samples were positive and 8 were negative among 50 positives by RBT-TAT. Therefore, it showed 89.4% of specificity and 84% of sensi-tivity. Through the current experiments, we could set up an ELISA based on the omp which has low cross-reaction and high immunogenicity and concluded that the omp could be a good material for accurate diagnosis of bovine brucellosis.

Comparative Studies on Serological Tests for Actinobacillus pleuropneumoniae Infection in Swine (돼지에서 Actinobacillus pleuropneumoniae의 혈청학적 진단법에 대한 비교연구)

  • 심항섭;우종태;조중현;전무형
    • Korean Journal of Veterinary Service
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    • v.17 no.2
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    • pp.95-113
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    • 1994
  • To establish an effective diagnostic measure for detection of the antibodies against Actinobacillus pleuropneumoniae, the methods for tube agglutination test (TAT), plate agglutination test (PAT), micro-agglutination test(MAT) and agar-gel immunodiffusion test(ID) were improved and standarized, and the comparative studies were carried out. The results obtained through the experiments were summarized as follows. 1. The rabbit hyperimmune sera to reference serotypes 1 to 6 were cross-tested with TAT, PAT, MAT and ID. In the homologous systems, the range of antibody titers in TAT was 80 to 640, showing the cross-reaction in serotypes 3, 4, 5 and 6. The range of antibody titers in PAT was 4 to 64, showing the cross-reaction in serotypes 3, 4, 5 and 6. In ID, the range of antigen titers was 8 to 32, and cross-reaction was observed in serotype 5. 2. The optimal concentration of antigen in PAT and MAT were 100mg /ml and 1.25mg /ml respectively. The most sensitive reaction in MAT was observed in 52$^{\circ}C$ for 18hrs. 3. In ID, the most promising antigen and the buffer for agar-gel were EDTA-treated antigen and 0.05M tris buffer (pH 7.2), respectively. 4. By the tests for 200 swine sera, it was found that the frequency of positive reaction were 203 in TAT, 240 in PAT and 163 in ID. 5. When compared the titers of TAT with those of MAT for 200 swine sera, MAT showed the higher titer than TAT being increased by relative correlation. Int was found that the titer for positive readings were 20 in TAT and 40 in MAT. 6. when compared the results of ID with those of TAT for 200 swine sera, all sera with TAT titer under 10 were negative in ID. Of the sera with TAT titer 20 and 40, 55.1% nd 91.8% were positive in ID, respectively. All sera with TAT titer above 80 were positive in ID. In comparison of ID and MAT, all sera with MAT titer under 20 were negative in ID. Of the sera with MAT titer 40 and 80, 24.7% and 93.9% were positive in ID, respectively. All sera with MAT titer over 160 showed positive in ID. 7. In conclusion, the established MAT showed high sensitivity but low specificity, wherease ID revealed low sensitivity but high specificity.

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Optimal Condition and Interspecific Cross-Reaction of H-Y Antibody Activity (H-Y항체활성의 최적조건과 종간교차반응)

  • ;H.S.Shim;J.B.Kim;H.Y.Park;K.S.Chung
    • Korean Journal of Animal Reproduction
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    • v.10 no.2
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    • pp.168-174
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    • 1986
  • These experiments were carried out to clarify the optimalconditions and interspecific cross reaction of H-Y antibody activity. H-Y antiserum was prepared in inbred SD female rats and Balb/c female mice by repeated immunization of rat newborn testis homogemate, rat and mouse spleen cells obtained from males of same strain. The activity of H-Y antibody in antiserum was tested by ELISA and biological tests. The cross reactivity of H-Y antibody was confirmed by culturing mouse and rabbit embryos in medium containing H-Y antibody and complement obtained from rat and guinea pig, respectively. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos in medium with different pH and complement concentration. The results obtained in these experiments were summarized as follows: 1. The formation rates of H-Y antibody in rats immunized with newborn testis and spleen cell were 40.0 and 50.0% respectively, and that in mouse immunized with spleen cell was 48.4%. 2. The activity of H-Y antibody was not affected by pH in range of 6.5 to 8.0, and the same was true for the relative concentration of complement to the H-Y antibody. 3. Minimum time needed for the activity of H-Y antibody was confirmed to be 0.5 to 1 hour and 24 to 48 hours respectively for the zona free embryos and intact embryos. 4. When mouse and rabbit embryos were treated with H-Y antibody obtained from rat, 46.4 and 54.8% of embryos were retarded or destroyed. From these results it could be said that H-Y antibody had strong interspecific cross reactivity.

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