• Title/Summary/Keyword: virus assay

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Epstein-Barr Virus Antibodies in Korean Mothers and Their Neonates (산모와 신생아의 Epstein Barr Virus 항체가에 관한 연구)

  • Shin, Young Kyoo;Eun, Baik Lin;Park, Sang Hee;Lim, Chae Seung;Kim, Young Sik
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.121-127
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    • 1998
  • Purposes : This study was performed to evaluate the seropositivities and levels of term pregnant women and their neonates, and the transplacental transfer rate of maternal Epstein-Barr Virus-specific IgG(VCA IgG and EBNA IgG) from term pregnant women to their neonates. Subjects and Methods : During Jan. 1, 1997 to Mar. 31. 1997, we collected the 42 pairs of sera from pregnant women and umbilical cord of their neonates in Korea University Ansan Hospital. The serum levels of VCA IgG and EBNA IgG were measured by the ELISA method. Results : 1) The seropositivities of VCA IgG were 100% in mothers and neonates. There was no statistical difference of mean VCA IgG levels between mothers and neonates. There was significant correlation of VCA IgG levels between maternal sera and neonatal umbilical cord sera(correlation coefficient r=0.5214, P<0.001). 2) The seropositivities of EBNA IgG were 100% in mothers and neonates. There was no significant difference of the mean EBNA IgG levels between mothers and neonates. There was significant correlation of EBNA IgG levels between maternal sera and neonatal umbilical cord sera (correlation coefficient r=0.7244, P<0.001). 3) There was no correlation between VCA IgG and EBNA IgG levels of maternal sera. Conclusion : Seropositivities of EBV CA IgG and EBNA IgG of term-pregnant women and their neonates were 100% and no significant differences of antibody levels were found in two groups. It seems that EBV Antibody levels in Korean mothers and neonates were high enough to protect primary EBV infection during early infancy.

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Seroepizootiological Study on Bovine Leucosis in Korea (한국(韓國)에서의 소백혈병(白血病)의 혈청역학적(血淸疫學的) 연구(硏究))

  • Jun, Moo-Hyung;Chung, Un-Ik;Lee, Chang-Ku;Baig, Soon-Yong;Lim, Chang-Hyeong
    • Korean Journal of Veterinary Research
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    • v.22 no.2
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    • pp.175-185
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    • 1982
  • Since bovine lymphosarcoma causes considerable economic loss to the dairy industry, seroepidemiological survey on bovine leucosis virus (BLV) was carried out for the dairy herds throughout the country to observe the epidemiological situation of the disease by using immunodiffusion test. Attempts were simultaneously made to detect bovine leucosis virus in the lymphocytes from BLV antibody-positive cattle by means of fluorescent antibody techniques, syncytium assay and electron microscopy. In immunodiffusion test for BLV antibody in 2003 heads of dairy cattle selected randomly from 164 herds, the prevalence of positive reactors by regions were 37.8% in Central, 27.2% in Honam (Southwest), 28.0% in Youngnam (Southeast) and 25.2% in Youngdong (East coast)and averaging 29.7%. By provinces, Chungcheong appeared the highest prevalence of BLV antibody carriers (41.8%), while Jeonbug revealed the lowest incidence rate (24.4%). When the results of serological studies were analyzed by age groups and the sizes of herds, the number of reactors increased gradually with the advance in the age of cattle and the herd size. The highest rate of BLV carriers was found in the ages between 6 and 8 years, and in the size of herds with 20 to 50 heads. One hundred and seventeen breeding bulls from the central regions were tested for BLV antibody. Four out of 70 bulls (5.7%) of Korean cattle and 14 out of 39 bulls (35.9%) of Holstein were reactive for BLV antigens. Of 164 dairy herds examined, 17 herds (10.4%) have no BLV antibody-positive cattle, while 42 herds (25.6%) were included in the range of 20 to 40% of the positive rate and 10 herds (6.1%) in the range of over 80% of the rate. When the lymphocytes from the BLV antibody carrying cattle were cultured in the presence of phytohemagglutinin and stained with FITC-conjugated sheep anti-BLV serum, 8 out of 11 cases (72.7%) of BLV positive cattle revealed specific fluorescence for BLV in the lymphocytes. In syncytium assay of the peripheral lymphocytes of the cattle, 5 out of 7 (71.4%) lymphocytes from BLV antibody carriers induced syncytia in the indicators of bovine embryonic splenic cells. The cultured lymphocytes were examined with an electron microscope to detect the BLV particles. Two out of 6 specimens (33.3%) from the reactors showed the typical type C virus with the size of 90 to 110 nm around microvilli and in intracytoplasmic vacuoles.

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Development of a Duplex RT-PCR Assay for the Simultaneous Detection and Discrimination of Avirulent and Virulent Newcastle Disease Virus (NDV) (뉴캣슬병 바이러스 검출 및 병원성 감별을 위한 Duplex RT-PCR법 개발)

  • Kim, Ji-Ye;Lee, Hyun-Jeong;Jang, Il;Lee, Hee-Soo;Yoon, Seung-Jun;Park, Ji-Sung;Seol, Jae-Goo;Kim, Seung-Han;Hong, Ji-Mu;Wang, Zillian;Liu, Hualei;Choi, Kang-Seuk
    • Korean Journal of Poultry Science
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    • v.44 no.2
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    • pp.93-102
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    • 2017
  • A duplex RT-PCR (dRT-PCR) assay was developed for the simultaneous detection and discrimination of non-virulent and virulent Newcastle disease virus (NDV) in a single PCR tube. Primers targeting the large polymerase protein (L) gene and the fusion protein (F) gene of NDV were designed to detect all NDVs (by common type PCR primers) and virulent NDVs (by pathotype PCR primers), respectively and evaluated experimentally with reference NDV strains and other poultry viral pathogens. PCR products of the expected size of 386 bp were amplified from all NDV samples whereas PCR products of the expected size of 229 bp were amplified from virulent NDV samples alone. Cross reaction was not observed with other avian viral pathogens. The detection limit of NDV by the dRT-PCR was estimated to be $10^3$ 50% egg infectious dose/0.1 mL. In the dRT-PCR using field isolates of NDV, the pathotype PCR primers detected specifically all of virulent field isolates of NDV from Malaysia, Pakistan and China whereas common type PCR primers detected 94.4% (51/54) of field isolates of NDV from China. Three Chinese NDV isolates with false negative result were non-virulent viruses. Our results indicate that the dRT-PCR might provide a rapid and simple tool for rapid simultaneous detection and discrimination of non-virulent and virulent NDVs. Therefore the developed dRT-PCR assay provides a powerful novel means for the rapid diagnosis of Newcastle disease.

The usefulness of skin test in evaluation of immunity to varicella (수두에 대한 면역력 평가에 있어서 피부 시험의 유용성)

  • Kim, Ji-Young;Lee, Hye Jeen;Kim, Me Jin;Kim, Young Ho;Jung, Ji A;Yang, Seung;Hwang, Il Tae;Lee, Hae Ran
    • Clinical and Experimental Pediatrics
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    • v.51 no.4
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    • pp.377-382
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    • 2008
  • Purpose : The aim of this study is to assess the usefulness of skin test by an inactivated, 1/50 diluted solution of attenuated varicella vaccine in evaluating the immune status to varicella. Methods : Total 41 subjects (22 males, 19 females, aged 1-32 years) were enrolled from July to August, 2005. Past medical history including varicella infection, varicella vaccination were investigated through questionnaires. The skin test solution was prepared from solution of attenuated varicella vaccine(Oka strain) which was inactivated by exposure to room temperature for 10 days and diluted at 1/50 with normal saline. Skin test was done by injecting 0.1 mL of the solution intradermally into the volar surface of the right forearm and sterile normal saline was used as a control on the left forearm. Positive reaction was defined when the transverse diameter of the induration was 5 mm or more. Serum varicella zoster virus specific IgG antibody test by ELISA (enzyme-linked immunosorbent assay) was done. Results : In adults, the sensitivity of the varicella zoster virus skin test compared to ELISA was 94.7% and the positive predictive value was 100%. In children, both the positive predictive value and specificity were 100% but the sensitivity and the negative predictive value were 50% and 30.7% respectively. Children showed smaller skin test reactivity compared to adults. Conclusion : The varicella zoster virus skin test using inactivated, 1/50 diluted solution of attenuated varicella vaccine was proved as one of the useful tools for evaluating the immunity and susceptibility of the varicella zoster virus.

Transgenic tobacco culture cells expressing spike protein gene of porcine epidemic diarrhea virus (돼지 유행성 설사병 바이러스 스파크 단백질 유전자 발현 형질전환 담배 배양세포)

  • Yang, Kyoung-Sil;Kim, Hyeon-Soo;Kwon, Suk-Yoon;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.87-94
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    • 2008
  • Porcine epidemic diarrhea virus (PEDV) is an infectious and highly contagious virus of swine. In order to develop the transgenic tobacco culture cells producing PEDV antigen protein, four vectors expressing PEDV spike protein (SP) gene under the control of a CaMV 35S promoter were constructed. Four fragments of the SP region of PEDV, SP1 (444 bp, 1487-1930 bp), SP2 (1.7 kb, 2300-3987 bp), SP3 (1.4 kb, 1559-2950 bp), and SP4 (2.6 kb, 9-2643 bp) were amplified by PCR and then C-MYC tag was fused to the end of each SP gene, respectively. These cassettes are inserted into the pCAMBIA2300 (named as 35S::SP1-M, 35S::SP2-M 35S::SP3-M, and 35S::SP4-M, respectively). Tobacco (cv. BY-2) cultured cells were transformed by co-cultivation with Agrobacterium tumefaciens harboring expression vector. We selected kanamycin-resistant calli and checked for the presence of the introduced SP gene using PCR, resulting 70% of them showed the foreign gene. We selected the lines with high-level expression of PEDV antigen protein based on dot blot analysis. Southern blot analysis confirmed that the PEDV SP gene was integrated into the genome of the tobacco cultured cells. Northern blot analysis showed that the introduced gene was highly expressed in transgenic cultured cells. Transgenic tobacco cultured cells-derived antigen induced immunogenicity in mice as determined by a plaque reduction neutralization assay. These results suggest that the vectors expressing PEDV spike protein gene in this study will be useful for the development of transgenic plants and cultured cells producing PEDV antigene protein.

Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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Characterization of canine coronaviruses isolated from the dogs with diarrhea in Korea (설사증 이환견(犬)으로 부터 분리(分離)한 canine coronavirus의 성상(性狀)에 관한 연구(硏究))

  • Lee, Byung-hyung;Jun, Moo-hyung;Park, Jong-hyeon;Hwang, Eui-kyung;Huh, Won
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.517-527
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    • 1994
  • An attempt was made to isolate a causative viral agents from the fecal specimens of the diseased dogs with the gastroenteritis symptoms. Two coronavirus-like agents were isolated by serial dilution end point method and plaque assay. The isolates were characterized in terms of cytopathology, antigenicity, replication, physicochemical and morphological properties. The results obtained through the experiment were as follows; 1. Among 7 fecal specimens collected from the dogs with enteric disease, 2(28.6%)coronavirus-like agents showing typical cytopathic effects of canine coronavirus were isolated, and designated as CCV D1 and CCV D2, respectively. 2. By the cross-neutralization test and indirect immunofluoresence antibody test, the isolates were antigenically indentified as the standard CCV. The viruses were replicated only in the cytoplasm of A-72 cells. 3. The isolates showed no haemagglutinating activity against the erythrocytes from 11 kinds of animals. 4. The electron microscopic observation for the isolates showed spherical and pleomorphic features, covered with club-shaped projections on the surface. The size of particles was ranged from 70 to 150nm. 5. In one-step growth curve for the isolates in A-72 cells, maximum titers of intracellular vius was $10^{4.6}$ $TCID_{50}/0.1ml$ at 46 hrs postinoculation(pi) of CCV Dl and $10^{4.4}$ $TCID_{50}/0.1ml$ at 34 hrs pi of CCV D2. The maximum titers of extracellular virus was $10^{5.5}$ $TCID_{50}/0.1ml$ at 58 hrs pi of CCV D1 and $10^{5.8}$ $TCID_{50}/0.1ml$ at 46 hrs pi of CCV D2. 6. In physicochemical property test, the isolates were very sensitive to choroform and were found to be RNA virus. The viruses was stable at pH 3.0 for 1 hr and at $22{^{\circ}C}$ for 5 hrs. However, infectivity titers reduced remarkably by treatment with $56{^{\circ}C}$ for 10min.

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Pattern of the Occurrence of Tomato yellow leaf curl virus on Cultivation (재배작형에 따른 토마토황화잎말림바이러스병의 발병양상)

  • Ko, Sug-Ju;Choi, Duck-Soo;Ma, Kyung-Cheol;Kim, Do-Ik;Kim, Mi-Kyeong;Choi, Hong-Soo;Kim, Hyun-Woo
    • Research in Plant Disease
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    • v.20 no.4
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    • pp.303-306
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    • 2014
  • Tomato yellow leaf curl virus (TYLCV) disease survey of tomato plants cultivated at Youngam, Changseong and Hwasun in Jeonnam Province was conducted in 2012 to 2013. To detect TYLCV in the field, PCR assay was performed using genomic DNAs extracted from tomato samples. According to cropping pattern, 23 and 59.4% of TYLCV infections were shown in semi-forcing and retarding cropping systems, respectively, and the frequencies of diseased plants were 0.4 and and 13.1% in the two cropping systems, respectively. Especially, TYLCV incidences in the susceptible varieties were 24.3 and 83.0% in semi-forcing and retarding cropping systems, respectively. Resistant varieties showed 9.1% infection in retarding cropping system but did not in semi-forcing cropping system. Also, TYLCV was occurred on early June after transplanting on April and at approximate 20 days after transplanting on August or September.

Difference in the Transcriptional Activity of the Interleukin-4 Promoter Haplotypes (Interleukin-4 유전자의 Promoter 일배체형에 따른 전사능의 차이)

  • Choi, Eun Hwa;Kim, Hee Sup;Chanock, Stephen J.;Lee, Hoan Jong
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.495-499
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    • 2005
  • Purpose : Interleukin-4(IL-4) is a critical component of the Th2 cytokine pathway and contributes to severity of respiratory syncytial virus(RSV) bronchiolitis. Previous studies observed an association between severe RSV bronchiolitis in Korean children with a common haplotype of the IL4 promoter. This study was performed to investigate functional differences of the variant IL4 promoter haplotypes. Methods : Genomic DNA was obtained from 20 children from 6 to 48 months of age in the Department of Pediatrics, Seoul National University Bundang Hospital. The IL4 promoter spanning an 1.2 kb region was amplified and haplotype was determined by cloning and the PHASE reconstruction. Transcriptional activity of Jurkat T cells which were transfected with each IL4 haplotype were analyzed by use of luciferase assay. Results : Three haplotypes of the IL4 promoter have been identified with the frequency of GCC(7 percent), TCC(17 percent), and TTT(76 percent). The TTT haplotype demonstrated the highest luciferase values in both unstimulated and PMA-stimulated Jurkat T cells. Increases in transcriptional activity compared to GCC have been shown in TTT(5.3 fold higher) followed by TCC(4.2 fold higher) in unstimulated Jurkat T cells. Conclusion : We provided evidence that increased transcriptional activity of the TTT haplotype of the IL4 promoter, which has previously been over-represented in Korean children with severe RSV bronchiolitis. Therefore, IL-4 could play a potential role in the pathogenesis of RSV infection, possibly via an altered transcriptional activity of the different IL4 haplotypes.

Environmental Control Accomodative to Ecosystem on the Prevention of Nuclear Polyhedrosis Virus Disease in the Silkworm. Bombyx mori (생태친화적(生態親和的) 사육환경(飼育環境) 제어방식(制御方式) 확립(確立)을 위한 누에 핵다각체병(核多角體病) 발생요인(發生要因) 분석(分析))

  • Han, Myung-Sae;Lim, Jong-Sung
    • Current Research on Agriculture and Life Sciences
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    • v.15
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    • pp.69-81
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    • 1997
  • For the prevention of worldwide prevalent disease of nuclear polyhedrosis virus (NPV), environmental conditions and their incidence of grasserie was investigated through 57 cases of silkworm rearing from the year of 1979 to 1993 in the countries of Korea, Japan, and Philippines. Relationship between the occurrence of NPV and environmental factors were also analysed from the aspect of causal pathogenesis. Unfavorable foactors related to the prevalence of NPV disease was reconfirmed by the assay of experimental rearing. Silkworms reared on mulberry leaves or artificial diet appeared similar result on the occurrence of grasserie. Disinfection by formalin and simple sweeping or washing was not significantly different on the occurrence of NPV disease. Following insufficient ventilation on the younger larvae. from the 1st to 3rd instar, the disease by NPV at the later stage was remarkably emphasized those insidence. An experimental rearing from 1993 to 1996 demonstrated the prevention of NPV disease by simple cleaning of sweeping under the condition of air forced ventilation, the customal practice of disinfection with formalin or any other chemical agents could be omissible.

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