• Title/Summary/Keyword: immunofluorescent antibodies

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Prevalence of Antibodies to Human Herpesvirus 8 in Children (소아의 항 Human Herpesvirus 8 항체 양성률)

  • Han, Tae Hee;Chung, Ju Young;Kim, Sang Woo
    • Pediatric Infection and Vaccine
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    • v.12 no.2
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    • pp.108-113
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    • 2005
  • Purpose : Human herpsevirus 8(HHV-8), a gamma herpsevirus, was initially identified from Kaposi sarcoma(KS) lesions and has been known to be associated with several malignancies including Kaposi sarcoma. HHV-8 seroprevalence is variable by different geographic areas and populations. The prevalence of HHV 8 infection in Korean children is unclear. So, we investigated the prevalence of HHV-8 specific antibodies in healthy children in Seoul, Korea. Methods : Sera were obtained from 112 children(age 1~15 years, 64 males and 48 females) who visited our hospital for routine health checkup and used for investigating sero-prevalence of anti-HHV-8 antibodies. An indirect immunofluorescent assay was used to detect the IgG antibodies to the lytic viral antigen(Biotrin, Dublin, Ireland). A peptide mix ELISA kit was used to detect the IgG antibodies to peptides specific for HHV-8 open reading frame (ORF)(Biotrin, Dublin, Ireland). Results : Of 112 children, 4 children younger than 6 years of age were seropositive to HHV-8[all 4(3.5%) were positive by IFA and 2(1.8%) were positive by ELISA]. Conclusion : These results suggest that the prevalence of antibody to HHV 8 in children in Korea is very low.

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Development and Characterization of Monoclonal Antibodies against Nucleoprotein for Diagnosis of Influenza A Virus

  • Nguyen, Hong Phuong;Kwak, Chaewon;Heo, Chang-Kyu;Cho, Eun Wie;Yang, Jihyun;Poo, Haryoung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.809-815
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    • 2018
  • Influenza, which is a highly contagious disease caused by the influenza A virus, continues to be a major health concern worldwide. Although the accurate and early diagnosis of influenza virus infection is important for controlling the spread of this disease and rapidly initiating antiviral therapy, the current influenza diagnostic kits are limited by their low sensitivity. In this study, we developed several new influenza nucleoprotein (NP)-specific monoclonal antibodies (mAbs) and compared their sensitivity and specificity of those with commercially available anti-NP mAbs. Three mAbs, designated M24.11, M34.3, and M34.33, exhibited higher reactivities to recombinant NPs and A/Puerto Rico/8/1934 (H1N1) viral lysates compared with the commercial mAbs, as assessed using enzyme-linked immunosorbent assays. M34.3 and M34.33 showed higher reactivities with A/California/04/09 (pandemic H1N1) and A/Philippines/2/82 (H3N2) viral lysates than the commercial mAbs. In contrast, M24.11 had marked reactivity with H3N2 but not with pandemic H1N1. Immunofluorescent confocal microscopy showed that the three mAbs effectively detected the presence of influenza virus in lung tissues of mice infected with A/Puerto Rico/8/1934. These results indicate that the newly developed M34.3 and M34.33 mAbs could be useful for the development of influenza diagnostics.

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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Seroepidemiologic Analysis of Acute Febrile Illness from Korea in 1996 (1996년 한국에서 발생한 열성질환에 대한 혈청역학적 연구)

  • Song, Jin-Won;Han, Sung-Hee;Baek, Luck-Ju;Lee, Yong-Ju;Song, Ki-Joon
    • The Journal of Korean Society of Virology
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    • v.28 no.4
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    • pp.377-382
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    • 1998
  • Hemorrhagic fever with renal syndrome (HFRS), scrub typhus, murine typhus and leptospirosis have been the principal acute febrile diseases in Korea. To evaluate the seroepidemiologic patterns of acute febrile illness, sera collected from 2,423 patients in 1996 were examined for antibodies against Hantaan virus, Orientia tsutsugamushi, Rickettsia typhi, and Borrelia burgdorferi by indirect immunofluorescent antibody technique (IFA) and macroscopic agglutination test for Leptospira interogans. Seropositive cases against Otsutsugamushi, Rickettsia typhi, Leptospira interogans and Hantaan virus were 192 (7.9%), 193 (8.0%), 12 (0.5%) and 324 (13.4%), respectively. Male was more affected in HFRS and murine typhus contrasting to scrub typhus and leptospirosis in female. Most positive cases occurred during October and November for scrub typhus, and during November and December for HFRS. These results showed similar patterns with previous epidemical data for recent couple of years, and possibly implied no significant changes occurred in ecologic situations for acute febrile diseases in Korea.

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Analysis of the Antigenic Expression Patterns of Herpes Simplex Virus Type 1 in BALB/c (BALB/c에서 Herpes simplex 1형 바이러스 항원 발현 양상에 따른 분석)

  • 고승석;조명환
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.62-66
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    • 2001
  • This study was performed to investigate antigenic expression patterns in the course of HSV-1 infection. In SDS-PAGE analysis, HSV-1 antigens were detected, and among them, antigens in the size of 39, 47, 63, 86, 101, 105, 135, 159, and 181 kDa appear to be expressed in the most dominant forms. BALB/c mice were infected with HSV-1 for 29 days and antigenic expression from HSV-1 was investigated by Western blot analysis using anti-HSV-1 sera collected every two days from BALB/c mice infected with HSV-1. Most of HSV-1 antigens appeared sporadically as the infection progressed. However, antigens in the sizes of 63kDa and 135kDa were expressed from day 1 and 3, respectively, and existed continuously during the course of infection for 29 days, suggesting that they are the most dominant antigens inducing immune response durign HSV-1 infection, and they could be the target antigens for the development of vaccines. The isotype levels of IgA, IgGl, and IgM increased till the 17 th day infection and then started to decrease. During this course. IgGl was the most dominant isotype. In an indirect immunofluorescent assay, antibodies exhibited surface binding to the Vero cell infected with HSV-1, demonstrating that HSV-1 antigens are expressed on the surface of Vero cells.

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Pathologic Studies in Piglets Naturally Infected with Porcine Reproductive and Respiratory Syndrome Virus (돼지 생식기 호흡기 증후군 바이러스 자연감염 예의 병리학적 연구)

  • Kim, Jae-Hoon;Hwang, Eui-Kyung;Kim, Yong-Joo;Sohn, Hyun-Joo
    • Korean Journal of Veterinary Pathology
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    • v.1 no.2
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    • pp.125-134
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    • 1997
  • Porcine Reproductive and Respiratory Syndrome Virus infection (PRRSV) was confirmed by serology histopathology immunohistochemistry and bacteriologic examination in young pigs. Four suckling and six weaned piglets submitted from three different farms showed coughing sneezing labored rapid abdominal respiration lethargy and anorexia. Grossly apical and cardiac lung lobes appeared mottled with pale to dark tan discoloration. Submandibular and bronchial lymph nodes were tan and enlarged. All piglets were seropositive for PRRSV antibodies by the indirect immunofluorescent antibody(IFA) test. Microscopically lung lesions were characterized by hyperplasia and hypertrophy of type 2 pneumocytes infiltration of mononuclear cells in alveolar intersitium accumulation of necrotic debris in alveolar spaces accompanied by proliferation of alveolar multinucleated syncytial cells. Using immunohistochemical technique PRRSV antigens were demonstrated in alveolar macrophages and type 2 pneumocytes in histologic lung tissue sections. Also PRRSV antigens were detected in brain lymph nodes spleen and heart. Additionally piglets showed nonsuppurative meningoencephalitis mandibular necrotic lymphadenopathy splenic atrophy and myocardial necrosis.

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A Study on Peripheral T Cell Subsets in Patients with Thyroid Tumors (갑상선 종양에 있어서 말초혈의 T 세포 Subset에 관한 연구)

  • Kim, Dong-Soo
    • The Korean Journal of Nuclear Medicine
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    • v.25 no.1
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    • pp.110-116
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    • 1991
  • To elucidate alteration of peripheral T cell subsets in thyroid tumors, the author enumerated T cell subsets in periphral blood by indirect immunofluorescent method, using monoclonal antibodies (CD3, CD4 and CD8) in 17 cases of thyroid cancer, 12 cases of thyroid adenoma, and 16 cases of adult healthy subjects as controls. Diagnoses were confirmed histopatologically in thyroid cancer and adenoma, and were established on the basis of commonly accepted clinical and biochemical criteria in Hashimoto's thyroiditis. The blood was drawn from veins of the patients and control subjects in Pusan National University Hospital during the period of January to October 1990. The results obtained were summarized as follow: 1) The percentage of CD3+ cells was significantly decreased in thyroid cancer as compared with healthy subjects. 2) The percentage of CD4+ cells was not different among thyroid cancer, thyroid adenoma, Hashimoto's thyroiditis and control subjects each other. 3) The percentage of CD8+ cells was significantly decreased in thyroid cancer as compared with adult healthy subjects, and tended to be decreased as compared with thyroid adenoma and Ha-shimoto's thyroiditis. 4) The CD/CD8 ratio was significantly increased in thyroid cancer as compared with control subjects, and tended to be increased as compared with thyroid adenoma and Hashimoto's thyroiditis. On the basis of the results, it can be suggested that the immunodysfunction may be due to decreased soppressor/cytotoxic T cells in thyroid cancer.

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Discrimination of Hantaviruses from the Tissues of Infected Hamsters to 5 Different Serotype Hantaviruses by Nested RT-PCR using Hantavirus Serotype Specific Primers (한타바이러스 혈청형 특이 Primer를 이용한 Nested RT-PCR 방법으로 5가지 혈청형 한타바이러스에 감염된 햄스터 조직에서 바이러스 검출)

  • Chu, Yong-Kyu;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.49-57
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    • 1997
  • We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

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

  • Kim, Hee-sun;Graham, Robert Ross;Lee, Jun-hak;Kang, Mun-il;Koh, Hong-bum
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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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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Association of BAF53 with Mitotic Chromosomes

  • Lee, Kiwon;Shim, Jae Hwan;Kang, Mi Jin;Kim, Ji Hye;Ahn, Jong-Seong;Yoo, Soon Ji;Kim Kwon, Yunhee;Kwon, Hyockman
    • Molecules and Cells
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    • v.24 no.2
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    • pp.288-293
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    • 2007
  • The conversion of mitotic chromosome into interphase chromatin consists of at least two separate processes, the decondensation of the mitotic chromosome and the formation of the higher-order structure of interphase chromatin. Previously, we showed that depletion of BAF53 led to the expansion of chromosome territories and decompaction of the chromatin, suggesting that BAF53 plays an essential role in the formation of higher-order chromatin structure. We report here that BAF53 is associated with mitotic chromosomes during mitosis. Immunostaining with two different anti-BAF53 antibodies gave strong signals around the DNA of mitotic preparations of NIH3T3 cells and mouse embryo fibroblasts (MEFs). The immunofluorescent signals were located on the surface of mitotic chromosomes prepared by metaphase spread. BAF53 was also found in the mitotic chromosome fraction of sucrose gradients. Association of BAF53 with mitotic chromosomes would allow its rapid activation on the chromatin upon exit from mitosis.