• 제목/요약/키워드: Indirect Immunofluorescence

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White spot syndrome virus (WSSV)에 대한 단클론 항체 생산 및 특성 분석 (Production and characterization of monoclonal antibodies against white spot syndrome virus (WSSV))

  • 공경희;김춘섭;김도형;김위식
    • 한국어병학회지
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    • 제35권2호
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    • pp.241-246
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    • 2022
  • White spot syndrome virus (WSSV) is a prevalent and virulent pathogen affecting cultured whiteleg shrimp (Litopenaeus vannamei) in Korea. In this study, seven monoclonal antibodies (mAbs) (10A12, 16C3, 17G4, 21G5, 22C4, 23B6 and 24G6) were produced by using purified WSSV. The reactivity of these mAbs was analysed by Western blot (WB), indirect immunofluorescence (IIF), and lateral flow immunochromatographic assay (LFIA). WB analysis demonstrated that three mAbs (17G4, 22C4, and 23B6) reacted specifically to VP28 with an approximate molecular weight of 24 kDa, mAb 16C3 reacted with approximately 17 kDa. IIF analysis demonstrated specific fluorescence signals on gill tissues of WSSV-infected shrimp, with five mAbs (10A12, 16C3, 22C4, 23B6, and 24G6), pleopods from WSSV-infected shrimp were used for LFIA, where, two mAbs (21G5 and 22C4) exhibited positive reaction. In conclusion, it can be inferred that the mAbs usage and specificity depends on the nature of assay used for diagnosis.

H-Y항체에 의한 생쥐초기배의 성판별에 관한 연구 II. 간접면역형광측정법에 의한 성판별 (Study on the sexing of preimplantation mouse embryo exposed to H-Y antisera II. Sexing of mouse embryos by immunofluorescence assay)

  • 양부근;장정순;김정익
    • 한국가축번식학회지
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    • 제12권1호
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    • pp.37-41
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    • 1988
  • These studies were carried out to examine the sex of preimplantation mouse embryo. For the investigation of sex-ration of mouse embryos, morula and blastocysts stage embryos treated with H-Y antiserum (10%, v/v) and FITC anti-mouse-IgG were divided into the positive and negative embryos. Positive and negative identified embryos were observed the viability according to the in vitro cultured and the sex ratio was also investigated by chromosomal analysis. The results obtained in these studies were summarized as follows: 1. Two hundred sixty-seven recovered embryos of morula or blastocyst stage were incubated in medium containing H-Y antiserum and FITC anti-mouse-IgG. Positively or negatively identified embryos were 139 and 128. This trend indicated the approximal sex ratio was 1:1. 2. Sex ratio was measured using the embryos treated with indirect immunofluorescence assay to examine the relationship between embryo developmental stage and sex ratio. Sex ratio of morula stage embryos was 45.2% positive and 54.8% negative, on the other hand, the ratio switched to 56.4% positive and 43.6% negative embryo in blastocyst stage. 3. Fourty-seven positive and 57 negative embryos were obtained out of 104 morula stage embryos treated with indirect immunofluorescence assay. Survived positive or negative embryos during in vitro culture were 42 and 49, respectively out of 47 and 57 embryos. 4. The numbers of negative and positive embryos were 171 and 92 out of 163 blastocyst embryos which were incubated in the medium containing H-Y antiserum and FITC anti-mouse-IgG. The result of karyotype test showed the successful rate of sexing embryo is positive and negative embryos was63.0% (58/92) and 62.0% (44/71). The final female to male ratio within 58 positive embryos was 22.7:77.6, and the ratio of the 44 negative embryos was 77.3:22.7.

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Expression of Human Cytomegalovirus Immediate Early US3 Gene in Human Fibroblast Cells

  • Lee, Gyu-Cheol;Lee, Chong-Kyo;Ahn, Jin-Hyun;Lee, Chan-Hee
    • Journal of Microbiology
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    • 제38권1호
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    • pp.24-30
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    • 2000
  • US3 gene is a member of the human cytomegalovirus (HCMV) immediate early gene. Although the precise functions of the US3 gene in HCMV replication and pathogenesis are not known, it has been reported to play a role in inhibiting major histocompatibility class I antigen presentation. For further knowledge of US3 gene expression, rabbit polyclonal antiserum of the US3 gene product was used for indirect immunofluorescence assay. In permissive human foreskin fibroblast (HFF) cells, US3 gene expression was detectable as crescent or half-moon shape in the perinuclear region at immediate early times after virus infection. HFF cells infected with mutant HCMV lacking US3 open reading frames were negative for US3 immunofluorescence assay. Double immunofluorescence assay using monoclonal antibody to gamma adaptin (specific for the Golgi complex) and rabbit anti-US3 antiserum revealed that US3 gene product could be localized to the Golgi complex. At later time after HCMV infection, US3 gene products were detected as globular aggregates in the cytosol. These aggregates were positive for gamma adaptin and stained with preimmune serum, suggesting a nonspecific reaction to the Golgi complex. Northern blot analysis revealed that transcription of US3 was observed only during immediate early times after virus infection (until 6 h postinfection). Therefore US3 gene expression appears to be confined to immediate early time and its gene products are localized to the Golgi complex as crescent shaped forms in the perinuclear cytoplasm.

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형광 현미경법 및 효소결합 면역협착법을 이용한 Bacteriodes gingivalis, Bacteriodes intermedius 및 Bacteriodes asaccharolyticus의 혈청학적 연구 (Serological study on the cross-reactivity of bacteroides gingillalis, bacteroides intermedius and bacteroides asaccharolyticus by indirect immunofluorescence and enzym-linked immunsorbent assay)

  • 정종평;이진용;이영희;정해원;정현주
    • Journal of Periodontal and Implant Science
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    • 제16권1호
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    • pp.100.1-100.1
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    • 1986
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The first virus isolation and partial characterization of equine herpesvirus-4 in a horse, South Korea

  • Choi, Eun-Jin;Lee, Hyun-Kyoung;Lee, Kyoung-Hyun;So, Byoung-Jae;Song, Jae-Young;Do, Jae-Chul;Yang, Seon-Joo;Lee, Hyun-Chul;Yang, Young-Jin
    • 한국동물위생학회지
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    • 제38권2호
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    • pp.141-144
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    • 2015
  • An equine herpesvirus-4 (EHV-4) was isolated in nasal swabs collected in a horse showing respiratory clinical signs. Equine dermis cells inoculated with the sample were observed with characteristic viral cytopathic effects after 3 days of postinoculation and the infected cells exhibited bright intracelluar fluorescence by indirect immunofluorescence assay. At the nucleotide level, the partial glycoprotein B gene of the Korean EHV-4 isolate (K001) had 99.9% identity to 1942 strain (GenBank No. M26171). To author's knowledge, the report describes the first isolation and partial characterization of EHV-4 in Korea. The virus can be used for further study of EHV-4.

배양 계배 신경아세포의 분화에 미치는 insulin의 영향 (Effect of Insulin on Differention of Chick Embryonic Neuroblasts Cultured in vitro)

  • 이창호;최덕영;박혜경;곽규봉;김혜선;정진하;하두봉
    • 한국동물학회지
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    • 제34권2호
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    • pp.209-216
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    • 1991
  • 신경세포의 분화에 미치는 insulin의 영향을 알아보기 위하여 계배의 단뇌 신경아세포를 serum-free defined medium에서 배양하였다. Immunofluorescence실험을 통하여 신경특이단백질인 MAP-2는 신경아세포의 세포체와 신경돌기에 존재하는 것으로 나타났다. 또한, 배양액내에 insulin의 농도를 증가하면 신경아세포의 신경돌기 형성이 증가할 뿐 아니라 MAP-2의 합성도 증가하였다. 따라서, 신경아세포의 형태적분화와 생화학적 분화는 서로 밀접하게 연관되어 일어나는 것으로 추측되며, insulin은 신경특이단백질의 합성을 촉진시킴으로써 신경아세포가 신경돌기를 형성하며 분화되어 가는데 결정적인 역할을 하는 것으로 보인다.

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Synthesis of New Uracil-5-Sulfonamide Derivatives and Immuno-Stimulatory Effect of a Chemically Modified Hemolymph of Biomphalaria alexandrina on Schistosoma mansoni Infected Mice

  • Fathalla, O.A.;Haiba, M.E.;Maghraby, A.S.
    • Archives of Pharmacal Research
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    • 제26권5호
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    • pp.358-366
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    • 2003
  • Some N-p-substituted phenyl uracil-5-sulphonamide derivatives have been synthesized to be evaluated as molluscicides against Biomphalaria alexandrina snails, the intermediate host of Schistosoma mansoni. Schistosoma mansoni infected mice were treated with hemolymph obtained from pre-treated Biomphalaria alexandrina snails with the products 4a, 10a, 10b and 4b or obtained from non-treated snails. The selection of the concentration based on the predetermined dose which caused mortality of less than 50% of snails/24 h. $LC_{50}$ of compounds 4a, 10a, 10b and 4b was 50, 100, 200 and 50 ppm respectively. The result showed that immuno-stimulatory effect of treated hemolymph with compounds 4a, 10a and 4b was related to significant protective effects (44.4, 34.6 and 50.4% reduction in worm burden respectively). In addition, mean total worm burdens were significantly reduced in non treated hemolymph by 33.8%. The effect of hemolymph obtained from treated or non treated snails on S. mansoni adult worms antigens was studied by indirect immunofluorescence technique using chronic mouse sera (CMS). The results indicated that there was a strong reaction with epitopes in gut epithelium, tubercles, teigument and subtegumental musculature of untreated and treated S. mansoni adult worms antigens. Therefore, treatment of hemolymph obtained from pre-treated snails with compounds 4a, 10a, and 4b can stimulate specific immune response and induce protective effects against S. mansoni infection.

신체 부위별 표피에서 밀착연접 단백질 중 오클루딘의 발현도 연구 (The Expression Pattern of the Tight Junction Protein Occludin in the Epidermal Context When Comparing Various Physical Samples)

  • 김지숙;장형석
    • 대한임상검사과학회지
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    • 제47권4호
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    • pp.267-272
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    • 2015
  • 밀착연접(tight junction)은 사람의 표피에서 관찰되는 세포간 이음(intercellular junction) 중 하나로서, 표피 과립세포층의 세포간격에 위치하여 인접한 세포들을 접합시키고 전해질과 수분의 이동을 조절하는 장벽기능을 비롯한 다양한 생물학적 기능을 수행한다. 이러한 밀착연접의 대표적 단백성분인 occludin은 그 발현도가 밀착연접의 표피-생물학적 기능성과 높은 상관관계가 있는 것으로도 알려져 있다. 본 연구에서는 신체 여러 부위의 정상피부에서 occludin의 발현도 또는 발현정도의 상이성 여부를 알아보기 위하여 occludin에 대한 항체를 이용한 간접 면역형광검사를 시행하여 그 결과를 비교 검토하고자 하였다. 신체 7개 부위별로 각 4개씩의 정상 피부조직을 채취하여 항 occludin 항체를 이용한 간접 면역형광검사를 시행한 결과, 양성반응을 보이는 최종 희석배수의 역수(reciprocal of end-point titer)로 표시한 부위별 occludin 항원의 발현도는 두피에서 300, 얼굴에서 600, 목에서 300, 팔에서 200, 배에서 600, 그리고 등과 다리에서는 모두 400으로 관찰되어 신체 부위별 occludin의 발현도에는 통계학적으로 유의한 차이가 있음을 알 수 있었다(p=0.001). 본 연구에서 관찰된 신체 부위별 occludin의 발현도의 차이는 밀착연접의 부위별 발현의 차등을 시사하는 소견으로서, 표피 피부장벽의 생물학적 기능의 가능한 부위별 차등성과도 관계가 있을 것으로 사료된다.

보르나 바이러스를 실험감염시킨 동물에서 항체검출에 대한 세포효소면역반응법의 평가에 대한 연구 (Evaluation of a cell enzyme-linked immunosorbent assay for the detection of Borna disease virus antibodies in experimentally infected animals)

  • 이두식
    • 대한수의학회지
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    • 제32권3호
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    • pp.377-380
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    • 1992
  • 보르나병 바이러스 혈청학적 진단법에 있어서 새로 개발된 세포효소면역법의 항체검출에 대한 평가를 위하여 지금까지 주로 사용되어진 간접형광항체법과 네 종류의 실험동물에서 서로 비교하였다. 모든 동물에서 간접형광항체법의 역가와 세포효소면역반응치 사이에 상관계수가 모두 0.8 이상으로 고도의 유의성이 인정되었고 두 진단법의 일치율은 한 희석단계 이하로서 재현성이 아주 좋았다. 세포효소면역반응법은 보르나병 바이러스의 혈청역학적 조사 및 병인기전 연구에 유용하게 이용될 수 있을 것으로 생각된다.

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Neospora caninum 간접형광항체진단법 개발 및 국내 가축에서의 항체가 조사 (Development of indirect fluorescent antibody test and the prevalence of the antibody titer for Neospora caninum of domestic animal in Korea)

  • 조영미;강승원;최은진;정우석;윤용덕;황우석
    • 대한수의학회지
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    • 제38권3호
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    • pp.595-599
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    • 1998
  • The present study was undertaken to develop the kit for indirect immunofluorescence antibody test(IFAT) and to investigate the prevalence of N caninum in cattle and dogs in Korea. The neo-antigen kit of IFAT developed in our laboratory was proved diagnostic efficacy by compared with the kit of veterinary medical research and development(VMRD). A survey of N caninum infections among cow and dogs in elevn areas of southern part of country was performed using a IFAT. The infection rate of 190 nationwide cattle was 8.4%(16/190) but was 75%(45/60) at P area in Chunnam province where the cattle showed the abortion repeatedly. Any of the dogs was not N caninum -positive in the Kangwon and Kyonggi areas.

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