• Title/Summary/Keyword: indirect immunoperoxidase technique

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Immunohistochemical identification of newcastle disease virus with indirect immunoperoxidase technique (Indirect Immunoperoxidase 법을 이용한 조직내 뉴켓슬병 바이러스 항원동정)

  • Nho, Whan-goog;Sur, Jung-hyang;Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.30 no.3
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    • pp.309-315
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    • 1990
  • The present experiment was done to identify newcastle disease virus(NDV) antigens in frozen sections of various oragns from experimentally NDV-infected with indirect immunoperoxidase method. Section were incubated with rabbit anti-NDV polyclonal as first antibody, followed by incubation with goat anti-rabbit or protein A peroxidase conjugate. Positive reactions were often detected in the epithelium of trachea and in the lymphocyte of spleen at 24 hours after virus inoculation. the viral antigen was localized mainly in the cytoplasm of infected cells. The method approved to be highly specific for the identification of NDV and allowed a precise localization of the viral antigens in infected cells.

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Immunohistochemical study on the antigenicity of each organ structure of Clonorchis sinensis (간흡충 충체의 부위별 항원성에 대한 면역조직화학적 연구)

  • Jin Kim;Jong-Yil Chai;Weon-Gyu Kho;Kyu-Hyuk Cho;Soon-Hyung Lee
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.21-30
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    • 1991
  • An immunohistochemical study was performed to demonstrate comparative antigenicity of each body structure of the liver fluke, Clonorchis sinensis, such as the digestive tract, reproductive organs, excretory system, tegument and suckers. Indirect immunoperoxidase technique was applied, rising formalin-fixed and paraffin-embedded sections of C. sinensis as the antigen. Pooled cat sera obtained 10 weeks after an experimental infection with C. sinensis and peroxidase-conjugated goat anti-cat IgG were used as the primary and secondary antibodies, respectively. The intensity of immunohistochemical stain was very sensitive upon the titers of the primary and secondary antibodies, and their optimum dilutions were found to be 1:1,000∼1:2,000 and 1:1,000, respectively. The intestinal epithelial cells, intestinal content and excretory bladder showed strong positive coloring reactions even at lower titer (1 : 2,000) of the primary antibody, whereas the uterine wall and eggs, vitelline glands, and male reproductive organs showed only weak positive reactions despite an increase in the antibody titer (1:1,000). On the other hand, the suckers, tegument, subtegumental cells and other parenchyme portions did not reveal any positive immunoperoxidase reaction at the same antibody titers. From the above results, it is highly suggested that the most potent antigenicity of C. sinensis occur from their excretory-secretory substances originated from the digestive and excretory organs.

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Application of immunohistochemical technique for diagnosis of porcine epidemic diarrhea (돼지 유행성 설사(Porcine Epidemic Diarrhea)의 진단을 위한 면역조직 화학적 기법의 응용)

  • Park, Nam-yong;Cho, Kyoung-oh
    • Korean Journal of Veterinary Research
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    • v.34 no.4
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    • pp.805-813
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    • 1994
  • Immunohistochemical study on the intestinal tissues obtained from the 21 pigs of the 14 terms in Korea in which the clinical and epidemiological features had indicated the possible outbreaks of porcine epidemic diarrhea(PED) was performed using the indirect immunofluorescence test and/or the immunoperoxidase method in order to detect PED viral antigens in the infected cells of the intestines, and histopathological features were described as well. By immunohistochemical analysis, PED viral antigens were detected in the epithelial cells covering the small intestinal villi and recognized slightly in the cells lining the colonic surface epithelium as well. Occasional fluorescence was also seen in a few intestinal crypt epithelium. On light microscopy, the piglets with PED showed marked villous atrophy and fusion, and severe enterocyte degeneration and desquamation. On the other hand, the older pigs more than 4 week old age was mild villous atrophy and fusion, severe villous epithelial cell proliferation, and moderate mononuclear cell infiltration.

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Immunohistochemieal study on the antigenicity of body compartments of Payugonimus westermani (폐흡충 충체 부위별 항원성에 대한 면역 조직화학적 연구)

  • Lee, Sun-Hyeong;Seong, Suk-Hwan;Chae, Jong-Il
    • Parasites, Hosts and Diseases
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    • v.27 no.2
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    • pp.109-118
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    • 1989
  • Production of circulating specific antibodies to the lung fluke (Paragenimus westermani) by its host is well known and used in various kinds of immunodiagnostic methods, However, it has not been well documented which compartments (or structures) of the lung fluke are most responsible for the production of specific antibodies. The present immunohistochemical study was undertaken to demonstrate the antigenicity of each body compartment of p. westermani such as suckers, tegument, spines, vitelline glands, intestine, reproductive organs(male and female), and eggs. Indiret immunoperoxidase(IP) stain technique was applied, using formalin-fked, paraffin- embedded lung tissues of P westermani-infected cats sectioned in 4 Um thickness as the antigen and cat antisera (11~20 weeks of infection) as the primary antibody. Peroxidase-conjugated goat anti-cat IgG was used as the secondary antibody and diaminobensidine(DAB) as the coloring agent. Strong yellow or yellowish brown staining was regarded positive. The primary and secondary antibody dilutions were made at 1 : 500~1 : 2, 000 and 1 : 200~1 : 500 respectively, and IP stain was repeated 10 times for each dilution. A consistent result obtained was that the intestinal epithelial border, intestinal content, vitelline glands, and eggs scattered around the worm capsule showed strong positive staining, while uterine eggs and some parenchymal portions showed weak positive reaction. On the other hand, the suckers, tegument, spines, subtegumental cells, cytoplasm of intestinal epithelial cells, male reproductive organs, and ovary revealed negative staining. The body compartments showing higher antigenicity were, in the decreasing order, the intestinal epithelial border, intestinal content, eggs in the worm capsule, vitelline glands, uterine eggs, and parenchymatous portions. The intestinal epithelial border and luminal contents revealed positive staining even at a few concentration of 1 : 4, 000 primary antibody(secondary ab., 1 : 200) whereas the parenchymatous portion showed positive reaction only at higher concentrations than 1'500 (secondary ab., 1 : 200). The results suggest that the specific antibody responses of the host to p. westermani occur most strongly upon the excretes from the intestinal epithelium of the worm and e99s Produced around the worm capsule,

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IMMUNOHISTOCHEMICAL STUDY ON LYMPHOCYTE DISTRIBUTION IN ENDODONTICALLY TREATED AND UNTREATED PERIAPICAL LESIONS (근관치료전과 후의 치근단 병소에서 임파구의 분포에 관한 면역조직화학적 연구)

  • Oh, Tae-Seok;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.11 no.1
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    • pp.63-75
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    • 1985
  • This study was designed to identify lymphocytes and to compare the lymphocyte distribution in endoodontically treated periapical lesions with that in endodontically untreated periapical lesions by way of immunohistochemical staining. Twenty-one human dental periapical lesions were obtained, frozened, serially sectioned to $4-5{\mu}$, and stained using the three-stage indirect immunoperoxidase technique and monoclonal antibodies for detecting the presence of B,T lymphocyte and T suppressor cell. Following results were obtained; 1. All of the examined periapical lesions had positive staining for B,T lymphocyte and T suppressor cell. 2. The concentration of T lymphocytes in 18 lesions diagnosed as periapical cyst and granuloma in both groups was greater than that of B lymphocytes and 2 periapical lesions identified as abscess in treated lesions had more positive B lymphocytes than positive T lymphocytes. 3. The average numbers of T,B lymphocytes and T suppressor cells in Endodontically treated lesions were lower than those of untreated lesions, but no statistically significant difference was noted. 4. When the distribution ratios of T lymphocytes to B lymphocytes and T suppressor cells to T lymphocytes were compared in Endodontically treated lesions by the histological aspects of the lesions and at the intervals of the duration after Endodontic treatment, a statistically significant change was not found. 5. The mean values of T lymphocytes, B lymphocytes and T suppressor cells in Endodontically treated lesions were markedly decreased in the specimens obtained at 3 month after Endodontic treatment, but no statistically significant difference was found.

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