• 제목/요약/키워드: Indirect Immunofluorescence

검색결과 90건 처리시간 0.023초

돼지 써코바이러스 2형의 진단을 위한 PCR법 적용 (Application of PCR for diagnosis of porcine circovirus type 2)

  • 박효선;이효상;나기복;이관복;강수정;문순화
    • 한국동물위생학회지
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    • 제29권1호
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    • pp.1-8
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    • 2006
  • Porcine circovirus (PCV) is a small, nonenveloped virus that contains a single-stranded circular DNA genome of about 1.76 kb and belongs to the family circoviridae. The PCV-2 has been incriminated as the cause of post-weaning multisystemic wasting syndrome (PMWS) , an emerging disease in pigs. In the present study, a PCR assay was applied to detect PCV-2 in tissue samples. The presence of PCV-2 antigen in the porcine tissues was confirmed by indirect immunofluorescence (IIF) with PCV-2 specific monoclonal antibodies. And then DNA extracted from PCV-2 positive tissues was used as a template. One oligonucleotide primer suitable for PCR was selected from a published PCV-2 sequence (Genbank). Amplified PCR product was detected the same fragment lengths of 416 bp as a control. Based on these results, it was suggested that the PCR is a simple and sensitive method for support diagnostic purposes.

Production and Characterization of Monoclonal Antibodies to Bovine Brain Succinic Semialdehyde Reductase

  • Park, E.Y.;Park, S.Y.;Jang, S.H.;Song, M.S.;Cho, S.W.;Park, S.Y.
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1995년도 춘계학술대회
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    • pp.72-72
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    • 1995
  • Monoclonal antibodies against bovine brain succinic semialdehyde reductase were produced and characterized. A total of nine monoclonal antibodies recognizing different epitopes of the enzyme were obtained, of which two inhibited the enzyme activity and three stained cytosol of rat spinal cord neurons as observed by indirect immunofluorescence microscopy. When unfractionated total proteins of bovine brain homogenate were seperated by gel electrophoresis and immunoblotted, the antibodies specifically recognized a single protein band of 34 kDa, which comigrates with purified bovine succinic semialdehyde reducatase Using the antisuccinic semialdehyde reductase antibodies as probes, we investigated the cross-reactivites of brain succinic semialdehyde reductases from some mammalian and an avian species. The immunoreactive bands on Western blots appeared to be the same in molecular mass-34 kDa-in all animal species tested, including humans. The result indicated that brain succinic semialdehyde reductase is distinct from other aldehyde reductases and that mammalian brains contain only one succinic semialdehyde reductase. Moreover, the enzymes among the species are imunologically very similar, although some properties of the enzymes reported previously were different from one another.

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국내 야생 집쥐(Rattus norvigicus)의 유행성 출혈열바이러스 감염증의 역학조사 (Epidemiological survey on Hantaan-related virus infection of wild rats(Rattus norvigicus) in Korea)

  • 김희선;;이준학;강문일;고홍범
    • 대한수의학회지
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    • 제33권2호
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    • pp.321-326
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    • 1993
  • Sixty eight wild rats were caught from Seoul, Kyonggi, Kangwon, Cholla, and Kyongsang provinces in Korea. All of them were confirmed in the same species, Rattus norvigicus. All of sera from wild rats were examined by indirect immunofluorescent test against Hantaan-related virus. Detection of anti-body to Hantaan-related virus were 4 out of 68 rats(5.9%), including 2 of 8(25.0%) rats in Kangwon and 2 of 20 rats(10.0%) in Kyongsang province. No difference of the sero-positive rates between sex of rats tested to Hantaan-related virus was recognized. According to age, four rats with antibodies(1:320~640) to Hantaan-related virus belonged one subadult, one young adult, one middle aged adult, and one old adult, respectively.

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체외생산된 소 배반포기배 ICM세포에서의 EGF-R 발현 (Expression of Epidermal Growth Factor-Receptor (EGF-R) on the Inner Cell Mass (ICM) of Bovine IVM/IVF/IVC Blastocyst)

  • 박세필;;정길생;김은영;윤산현;임진호
    • 한국가축번식학회지
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    • 제21권1호
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    • pp.39-46
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    • 1997
  • 본 연구는 체외생산된 소 배반포기배의 inner cell mass (ICM) 세포에서 epidermal growth factor-receptor (EGF-R)의 발현 유무를 immunosurgery와 indirect immunofluorescence (간접 면역 형광방법)을 이용하여 조사하고자 실시하였다. 본 실험에 사용된 ICM 세포는 체외수정 후 7∼8일째에 회수된 소 배반포기배로부터 immunosurgery 방법을 실시하여 얻어졌으며, 회수된 ICM세포는 live/dead 염색방법을 통한 생사 유무와 EGF-R 발현 유무 조사에 공시되었다. 특히, 배반포기배에 대한 immunosurgery를 위해 trophectoderm 세포에 대한 rabbit anti-bovine trophectoderm cell antibody (RABTE)를 제조하여 사용하였다. 결과를 요약하면 다음과 같다. ICM세포의 회수율은 RABTE와 guinea pig serum (complement)에 각각 15∼30분과 15∼60분동안 처리했을 경우 16.7∼74.2%였으며, 또한 처리시간이 각각 30분과 30분일 때 가장 높은 회수율(74.2%)을 얻었다. Immunosurgery 후 얻어진 ICM세포의 생존 유무를 조사하기 위해 live/dead 염색 방법을 이용하였던바, ICM세포의 생존율은 complement가 60분 처리된 군(69.3%)을 제외한 모든 처리군에서 84.0∼91.6%의 높은 생존율을 나타냈다. 또한, 회수된 ICM세포에 대한 EGF-R의 존재를 확인하였다. 따라서, ICM세포에서의 EGF-R의 발현은 인위적으로 첨가된 EGF의 이용 가능성을 높임으로서 체외에서의 착상전 배 발달을 증진시킬 수 있을 것으로 사료된다.

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국내 양돈장에서 분리한 G9형 돼지로타바이러스의 특성 (Characterization of G9 genotype porcine rotavirus isolated in Korea)

  • 왕쥔후이;이승철;강신영
    • 한국동물위생학회지
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    • 제35권2호
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    • pp.83-90
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    • 2012
  • Porcine rotaviruses are the most common causes of viral gastroenteritis in piglets around the world. The major G genotypes of porcine rotaviruses causing diarrhea were G4, G5 and G11 genotypes. Recently, G9 genotype rotaviruses were problemed at swine farms and frequently recognized from diarrheic piglets. In this study, a porcine rotavirus (PoRV-1) was isolated from piglet showing diarrhea using MA104 cells and confirmed as rotavirus by electron microscopy, genomic RNA electropherotyping and indirect immunofluorescence antibody tests. The nucleotide sequence of the VP7 gene of PoRV-1 was determined and compared with those of other genotype rotavirus strains from other parts of the world. Also, the nucleotide sequences of VP4, VP6 and NSP4 genes of PoRV-1 were determined and compared with those of other rotavirus strains from other countries. The results showed that the PoRV-1 isolate belonged to the G9 genotype and the P, I and E genotypes of PoRV-1 were P[23], I5 and E1, respectively. The Korean G9 PoRV-1 isolate and its nucleotide sequence data would be usefully used for the development of porcine rotavirus vaccines in near future.

이유자돈 전신소모성증후군 이환 자돈에서의 바이러스성 원인체 검색 및 porcine circovirus 2 분리동정 (Detection of viral pathogens and isolation of porcine circovirus 2 from postweaning multisystemic wasting syndrome-affected piglets)

  • 박최규;김현수
    • 대한수의학회지
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    • 제44권4호
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    • pp.561-569
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    • 2004
  • To detect viral agents and isolate porcine circovirus 2 (PCV2), 60 samples of lung and lymph node were collected from 5 to 12 week-old pigs that had showed clinical signs of postweaning multisystemic wasting syndrome (PMWS). Polymerase chain reactions (PCRs) were conducted to identify the viral pathogens including PCV1, PCV2, porcine parvovirus (PPV) and porcine reproductive and respiratory syndrome virus (PRRSV) that have been considered to be the causal agents of PMWS. Among 60 samples, PCV 2 was detected from 57 samples but no PCV 1 was detected. PRRSV and/or PPV were also detected from 27 (47.4%) samples and 1 (1.8%) sample of these 57 PCV 2-positive samples, respectively. Tissue homogenates were inoculated onto PCV-free PK-15 cell monolayers. Seven isolates were confirmed as PCV 2 by multiplex PCR, indirect immunofluorescence assay, and transmissible electron microscopy. These date suggest that PRRSV is a major cofactors causing PMWS in pigs that were infected with PCV2 in Korea.

Quantification of Immunoglobulin G against Trypanosoma cruzi in Individuals with Chronic Chagas Disease Treated with Nifurtimox and Evaluated in Prolonged Follow-Up

  • Munoz, Gabriela;Vergara, Camilo;Martinez, Gabriela;Apt, Werner;Zulantay, Ines
    • Parasites, Hosts and Diseases
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    • 제57권1호
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    • pp.39-41
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    • 2019
  • In the indeterminate chronic period of Chagas disease (ChD) the treatment has not been conclusive, because the serological negativization requires many years. This study aims to evaluate the efficacy of nifurtimox (NF) in the treatment of chronic ChD in prolonged follow-up by serological techniques of indirect immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (ELISA) IgG comparing 2 groups of patients, treated and non treated. Mann-Whitney test was performed for ELISA and IFA, with significant difference between the groups (P<0.05). IgG levels were lower in individuals treated compared with untreated patients, indicating chemotherapeutic efficacy in prolonged follow-up.

야외 송아지 설사변에서 작은와포자충 검출에 대한 평가 (Evaluation for detection of Cryptosporidium oocysts in diarrheal feces of calves)

  • 위성환;주후돈;강영배
    • Parasites, Hosts and Diseases
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    • 제34권2호
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    • pp.121-126
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    • 1996
  • 작은와포자충이 송아지에서 설사에 직접적으로 영향을 미치는지를 알아보고자 야외에서 설사를 발현하는 4개월 이하의 송아지 분변 총 201건를 수거하여 DMSO-modified acid fast 염색법 (MAFS), commercial kit(Meridian Diagnostics, Cincinnati, Ohio) 그리고 본 실험실에서 만 든 단세포군항체(C6)를 이용한 간접형광항체법(IFA)으로 작은와포자충의 검출을 시도하였다. 작 은와포자충의 검출은 MAFS에서 29건(14.4%) commercial kit와 IFA에서 각각 23건씩(11.4%) 검출되었다. 작은와포자충 오오시스트의 검출에 사응된 진단기법간의 진단일치율을 비교 하였던 바 msS를 기준으로 하였을 때 IFA는 민감도 69%. 특이성 98%였다. Commercial kit와 IFA진단법에서는 양성 22건, 음성 177건이 서로 일치하였으며. commercial kit를 기준으로 할 때 IFA의 민감도는 96%, 특이성 99% 고리고 진단일치에 대한 기대치는 99%였다. 따라서 본 실험에 사용된 IFA는 commercialkit와 일치율이 높아 진단목적으로 사용될 수 있을 것으로 생각된다. 한편, IFA에서 작은와포자충이 검출된 23건의 분변에 대해 분변 g당 작은와포자충 오오시스트의 수(OPG)를 측정하였다. 14건이 $10^3$ 이하의 OPG수준을 나타냈으나 $10^5$ 이상의 OPG도 4건이었다. 작은와포자충에 감염되어 설사가 발현될 때의 OPG의 수준이 $10^5$ 이상임을 감안해 보면, 이번 조사된 송아지 설사변 201건 중 4건(5%)은 작은와포자충에 의한 설사로 분석되었다. 이러한 결과를 종합해보면, 국내메서 발생되는 송아지 설사 원인중 약 5%는 설사와 관련되는 다른 원인체와 관계없이 작은와포자충 단독감염만으로도 임상적으로 설사가 발현되고 있는 것으로 판단되었다.

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국내분리(國內分離) Aujeszky's disease virus의 실험적(實驗的) 감염자돈(感染仔豚)의 병리조직학적(病理組織學的) 관찰(觀察) (Histopathological observation on the piglets experimentally infected with Aujeszky's disease virus isolated in Korea)

  • 조우영;조성환
    • 대한수의학회지
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    • 제30권1호
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    • pp.93-102
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    • 1990
  • Thirty-day-old piglets were intranasally or subcutaneously inoculated with 2ml of Aujeszky's disease virus, NYJ-1 strain, at the titer of $10^{6.75}$ $TCID_{50}/0.1ml$, that was isolated from the diseased piglets in Korea, and histopathological studies were performed to elucidate the pathognomonic characters of the isolate. Results obtained through the experiments were as follows: 1. Major clinical signs on the 2nd and 3rd days post inoculation (p.i.) were fever, anorexia and dyspnea. On the 6th and 7th days p.i., nervous signs, severe dyspnea and salivation were observed in the group of intranasal inoculation, and one out of 3 piglets in this group died on the 7th day p.i.. General signs were more severe in the group of intranasal inoculation than the group of subcntaneous injection. Between the 8th and l0th days p.i., the signs subsided and the piglets were completely recovered from the illness. 2. Hematologically, most of the inoculated pigs showed a mild lymphocytopenia on the 5th and 6th days p.i.. 3. By necropsy, swelling and hemorrhagic lesions were observed in tonsil, central nervous system and lung. No specific changes were grossly found in other parenchymatous organs. 4. In histopathological study, degeneration and necrosis of nervous cells, non-suppurative meningoencephalitis, diffuse or focal gliosis, perivascular cutting and degeneration of ganglion cells were observed in central nervous system, and swelling and hemorrhagic changes were shown in the tissues of liver, lung and lymph nodes. 5. By indirect immunofluorescence antibody assay using ADV-monoclonal antibody, specific ADV antigens were detected in the tissues of tonsil, brain and spleen of the succumbed piglet. However, in the experimentally slaughtered piglets, the specific reactions were noted only in the tonsils.

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Inhibition of Herpesvirus-6B RNA Replication by Short Interference RNAs

  • Yoon, Jong-Sub;Kim, Sun-Hwa;Shin, Min-Chul;Lee, Dong-Gun;Hong, Seong-Karp;Jung, Yong-Tae;Khang, In-Gu;Shin, Wan-Shik;Kim, Chun-Choo;Paik, Soon-Young
    • BMB Reports
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    • 제37권3호
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    • pp.383-385
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    • 2004
  • RNA interference (RNAi) is a process of sequence-specific gene silencing, which is initiated by double-stranded RNA (dsRNA). RNAi may also serve as an antiviral system in vertebrates. This study describes the inhibition of herpesvirus-6B (HHV-6B) replication by short interference RNAs (siRNAs) that are targeted to the U38 sequence that encodes DNA polymerase. When virus-infected SupT1 cells were treated by siRNA, these cells blocked the cytopathic effect (CPE) and detected the HHV-6B antibody-negative in indirect immunofluorescence assays (IFA). Our result suggests that RNAi can efficiently block Herpesvirus-6B replication.