• Title/Summary/Keyword: immunofluorescent assay

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Distribution of Waterborne Enteric Viruses in Raw Water and Tap Water in Busan Metropolitan City (부산시 상수원수와 수돗물에서의 수인성 장관계 바이러스 분포조사)

  • 박홍기;정은영;이유정;정종문;최동훈;손희종;권기원;홍용기
    • Journal of Life Science
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    • v.13 no.2
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    • pp.197-205
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    • 2003
  • We detested waterborne enteric viruses from the raw water and tap water in Busan metropolitan city by the total culturable virus assay of EPA standard method. According to the results of survey from July 2001 to November 2002, thirteen out of twenty one in raw water samples were positive (61.9%) for enteric viruses and all of the treated water and tap water samples were negative. The enteric viruses in raw water were mainly distributed through the summer to the earl y winter, suggesting the seasonal characteristics of virus distribution in water The titer of enteric viruses per 100 liters of the raw water was ranged from 1.92 to 9.70 MPN by TCVA-MPN program. The isolated viruses were identified as either human poliovirus type 1 or enteroviruses by the immunofluorescent assay.

Comparison between indirect immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to porcine reproductive and respiratory syndrome virus(PRRSV) (돼지 생식기호흡기증후군 바이러스항체 검색에 있어 간접형광항체법(IFA) 과 효소면역법(ELISA)의 진단효율 비교)

  • Park, Choi-kyu;Lyoo, Young-soo;Lee, Chang-hee;Jung, Jong-wook
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.314-318
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    • 1998
  • An establishment of effective control measures to PRRSV infection in swine industry depends on a sensitive and specific diagnosis to detect either viral antigen and/or antibodies to PRRSV. Several diagnostic methods are available to detect antibodies against PRRSV, including IPMA, IFA and ELISA tests have been successfully developed. Sensitivity of the indirect immunofluorescent assay in MA-104 cells using Korean field isolate PL96-1 was superior to that of VR-2332 and field isolate PL96-2. Sensitivity and specificity of the IFA test with PL96-1 were comparable to those of commercial ELISA test kit but ELISA test was more sensitive for the detection of declining antibodies to PRRSV in finishing pigs. In this study we concluded that IFA and ELISA test could be utilized to detect antibodies to PRRSV and the results generated from these two tests were comparable and there were no significant difference between these two tests.

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Comparative evaluation of indirect immunofluorescent antibody test with enzyme-linked immunosorbent assay in serodiagnosis of human neurocysticercosls (뇌낭미충증의 혈청학적 진단에 있어서 간접 형광항체 반응 및 효소연결성 면역흡착 검사의 비교 평가)

  • Eom, Gi-Seon;Jo, Seung-Yeol;Im, Han-Jong
    • Parasites, Hosts and Diseases
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    • v.26 no.1
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    • pp.27-32
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    • 1988
  • The applicability of indirect immunoftuorescent antibody test (IFAT) was compared with enzyme-linked immunosorbent assay (ELISA) in sera from 163 cases of confirmed neurocysticercosis, 101 other neurologic and parasitic diseases and 100 normal controls. As antigen, frozen sections of a Taenia solium metacestode from a human brain was used in IFAT and cystic fluid was used in ELISA. For the detection of specific IgG antibody, IFAT was equally sensitive (89.6%) and specific (85.1%) as ELISA. The antibody titers by IFAT were correspondingly increased with mean absorbance of ELISA. The corresponding rate of positivity in the two techniques was 90.8%. Except for the difficulty in detecting antibodies in cerebrospinal fluid (CSF), IFAT was concluded to be very useful for the serodiagnosis of human neurocysticercosis.

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ALCAM is a Novel Cytoplasmic Membrane Protein in TNF-α Stimulated Invasive Cholangiocarcinoma Cells

  • Adisakwattana, Poom;Suwandittakul, Nantana;Petmitr, Songsak;Wongkham, Sopit;Sangvanich, Polkit;Reamtong, Onrapak
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3849-3856
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    • 2015
  • Background: Cholangiocarcinoma (CCA), or bile duct cancer, is incurable with a high mortality rate due to a lack of effective early diagnosis and treatment. Identifying cytoplasmic membrane proteins of invasive CCA that facilitate cancer progression would contribute toward the development of novel tumor markers and effective chemotherapy. Materials and Methods: An invasive CCA cell line (KKU-100) was stimulated using TNF-${\alpha}$ and then biotinylated and purified for mass spectrometry analysis. Novel proteins expressed were selected and their mRNAs expression levels were determined by real-time RT-PCR. In addition, the expression of ALCAM was selected for further observation by Western blot analysis, immunofluorescent imaging, and antibody neutralization assay. Results: After comparing the proteomics profile of TNF-${\alpha}$ induced invasive with non-treated control cells, over-expression of seven novel proteins was observed in the cytoplasmic membrane of TNF-${\alpha}$ stimulated CCA cells. Among these, ALCAM is a novel candidate which showed significant higher mRNA- and protein levels. Immunofluorescent assay also supported that ALCAM was expressed on the cell membrane of the cancer, with increasing intensity associated with TNF-${\alpha}$. Conclusions: This study indicated that ALCAM may be a novel protein candidate expressed on cytoplasmic membranes of invasive CCA cells that could be used as a biomarker for development of diagnosis, prognosis, and drug or antibody-based targeted therapies in the future.

Canine distemper outbreak in a zoo (동물원에서 집단 발생한 개 디스템퍼 감염증)

  • Hur, Kwon;Bae, Ji-Seon;Choi, Jae-Hoon;Shin, Nam-Sik;Lee, Ki-Whan;Kwon, Soo-Wahn;Kim, Dae-Yong
    • Korean Journal of Veterinary Pathology
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    • v.2 no.2
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    • pp.139-145
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    • 1998
  • A total of 5 animals including 3 raccoons, 1 badger, and 1 fennec fox kept in outdoor exhibits at the Everland Zoological Gardens showed depression, anorexia, dyspnea, serous oculonasal discharge, diarrhea, and convulsions. All the affected animals died within 10 days after the onset of clinical signs. This outbreak lasted about 4 months. On necropsy, major gross lesions were confined to the lungs. Red to grey sublobular to lobular consolidations with various sized tan to reddish spots were observed in the lungs. Histopathologically, the pulmonary lesions were characterized by acute to subacute bronchointerstitial pneumonia with secondary bacterial or adenoviral infections. Intracytoplasmic eosinophilic inclusion bodies compatible with canine distemper virus (CDV) were found in the lung, urinary bladder, kidney, intrahepatic bile duct, stomach, small and large intestines. Multifocal areas of severe demyelination and accumulation of gitter cells or nonsuppurative inflammation were seen in the brains of 2 raccoons. CDV -specific antigens were demonstrated in the lung sections on immunofluorescent assay. The present report describes an outbreak of CDV infection in a zoo and indicates the range of susceptible zoo animal species.

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Seroprevalence and epidemiological analysis of porcine reproductive and respiratory syndrome virus in Korea (돼지 생식기호흡증후군 바이러스의 항체분포 및 역학조사)

  • Park, Choi-kyu;Chang, Chung-ho;Kang, Yung-bae;Lee, Chang-hee;Lyoo, Young-soo;Kim, Hyun-soo
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.111-117
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    • 1999
  • A nation wide sero-epidemiological survey of porcine reproductive and respiratory syndrome(PRRS) was carried out to analyze the current status of the PRRS virus infections in the field using the indirect immunofluorescent antibody assay(IFA) with the field isolate PL96-1. Since the first report of the antibody detection to PRRSV in 1993, the prevalence of seropositive pigs has increased dramatically and the data indicate that over 21% of the pigs and around 60% of the farms showed seropositives to the PRRS virus. A slightly higher positive rate was recognized in breeders than fattenings and it might be due to the higher age at the time of testings. No significant regional differences were detected in the sero-epidemiological survey. Higher sero-positive rate in growers indicates that PRRSV infection in the field was common after weaning(around 40 days). However, the number of seropositive pigs were declined in fattening pigs. Sows showed around 26% of sero-positive rate that there is a higher chance of continuous virus circulation in the infected farms. Low rate of sero-positivity in boars(9.8%) implies that there is high demand in proper control measures to prevent virus spreading through breeding procedures such as natural or artificial insemination. Therefore it was concluded that PRRSV infection in domestic swine herds is endemic and the positive rate and economic loses will be increased by spontaneous infections in naive farms.

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Vascular Endothelial Growth Factor Effect on Notch 1 Expression and Proliferation of Fibroblast (혈관내피성장인자의 섬유아세포 증식과 Notch 1 발현에 대한 영향)

  • Koh, Sung-Hoon
    • Archives of Plastic Surgery
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    • v.37 no.1
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    • pp.7-11
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    • 2010
  • Purpose: Vascular endothelial growth factor (VEGF) is known as a growth factor of endothelium and fibroblast. The purpose is to know the VEGF effects on fibroblast proliferation and fibroblast's notch receptor expression. Methods: CCD-986sk fibroblast was purchased from the Korean Cell Bank and was used in XTT assay for proliferation and wound healing assay for migration. Immunofluorescent (IF) staining and western blotting were used in testing notch expression of fibroblast. Semiquantitative RT-PCR was used in checking notch 1 mRNA production by fibroblast. Student-t test was used for analyzing results. Results: Cell proliferation assay using XTT showed significant higher proliferation in VEGF treated fibroblast, $2.324{\pm}0.0026$ vs. $2.463{\pm}0.017$ (p=0.002). Wound healing assay showed longer migration in VEGF treated fibroblast (p=0.062). The fluorescence was brighter in VEGF treated cells of notch 1 IF staining. Notch 1 expressions and mRNA productions increased more in VEGF treated cells. Conclusion: VEGF stimulates fibroblast to proliferate, migrate and to express Notch 1 simultaneously. Notch receptor could be related to VEGF mediated wound healing.

Dot Blot Assay for Screening of Anti-hantavirus Antibodies by Using Nucleocapsid Protein of Hantaan Virus (한탄바이러스 핵단백질을 이용한 항 한타바이러스 항체 검색용 Dot Blot Assay)

  • Cho, Hae-Wol;Chung, Yeun-Jun;Kim, Chung-Lim;Ban, Sang-Ja;Nam, Jae-Hwan;Lee, Hyeong-Woo;Lee, Yoo-Jin;Kim, Eun-Jung
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.59-65
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    • 1996
  • For easy and rapid screening of hemorrhagic fever with renal syndrome (HFRS) without any laboratory equipment, dot blot enzyme immunoassay was developed and tried to detect anti-hantavirus antibodies. The nucleocapsid protein of Hantaan virus was isolated by affinity chromatography and used for making the dot strip. 28 of 29 Hantaan virus infected sera showed positive signals and 21 of 22 HFRS negative sera showed no positive signals. Anti-Seoul virus monoclonal antibody also exibited positive signal but the intensity of colorization was approximately 5 fold less than that of anti-Hantaan monoclonal antibody. The sensitivity of dot blot assay was equal or superior to indirect immunofluorescent assay (IFA) or ELISA test. Overall, the screening results with dot blot assay showed 92.2 % of concordance with IFA or ELISA test. This results suggests that dot blot assay could be applied a tool for easy and rapid screening of HFRS.

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Production of Monoclonal Antibodies by Hybridomas Sensitized to Sporozoites of Cryptosporidium parvum (Cryptosporidium parvum Sporozoites 에 감작된 Hybridomas 에서의 Monoclonal Antibody 생산)

  • Cho, Myung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.494-498
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    • 1989
  • Hybridoma cell lines, which secrete monoclonal antibodies (mAbs) against the surface antigens of Cryptosporidium parvum Sporozoites, were produced by fusing spleen cells of C. parvum Sporozoite-immunized mice with P3-X63-Ag8 myeloma cells. Two cloned antibody-secreting cell lines, Kor1 and Ea2, were established and produced IgG1 and IgG2a antibodies, respectively. Percoll-purified sporozoites were solubilized and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Western blot assay demonstrates that an antigen of 20-kDa was bound by monoclonals. By indirect immunofluorescence microscopy, mAb exhibited uniform binding to the sporozoite surface.

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The serological studies on infectious bursal disease (전염성 F낭병에 대한 혈청학적 연구)

  • 정영미;서석열;도홍기;조정곤;노수일
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.271-279
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    • 2000
  • This study was carried out to provide the fundamental information for development of proper vaccination program against infectious bursal disease(IBD) to the local chicken farms. The antigen detection was peformed from 8 samples of bursa of Fabricius with agar gel precipitation(AGP) and indirect immunofluorescent assay(IFA), And also, the antibodies in serum samples were detected by the various serological methods such as commercial ELISA assay, AGP and virus neutralization(VN) test. 1. The antigen detection rates were 25% for AGP which is 2 out of 8 farms and 10 out of 40 bursas, and 25% which Is 2 out of 8 farms and 20% 8 out of 40 bursas for IFA, respectively. 2. The mean titer of maternal antibody (>3,000) existed until 10 days of the age with ELISA-GMT. 3. The antibody positive rates which are over 80% showed until 5 days of the age with ELISA and at 10 days of the age with AGP except one, but none of them showed from 1 day of the age. This report came to conclusions that both the protective maternal antibody titers and the antigen positive rates were significant until at the 10 days of the age.

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