• Title/Summary/Keyword: virus assay

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Seroprevalence of Swine Influenza Viruses H1N1 and H3N2 in Gyeongbuk Province, Korea (경북지역 돼지인플루엔자 바이러스(H1N1, H3N2) 항체조사)

  • Chae, Tae-Chul;Kim, Seong-Guk;Cho, Kwang-Hyun;Eo, Kyung-Yeon;Kwon, Oh-Deog
    • Journal of Veterinary Clinics
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    • v.31 no.4
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    • pp.293-297
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    • 2014
  • Swine influenza is an acute respiratory disease prevalent in pig-growing areas worldwide. In total, 518 gilt and sow serum samples and 14 litters (66 samples) of aborted fetuses from 37 farms (average of 14 serum samples per farm) in Gyeongbuk Province were collected between September 2010 and May 2011. All samples were examined for antibodies to swine influenza virus (SIV) H1N1 and H3N2 using enzyme-linked immunosorbent assay (ELISA). The seropositive rates of gilt and sows were 59.8% (310/518) for SIV H1N1, 78.8% (408/518) for H3N2, and 55.6% (288/518) for both subtypes tested. The rate of aborted fetuses was 13.6% (9/66) for H1N1, 9.1% (6/66) for H3N2, and 9.1% for both subtypes. The seroprevalence for H1N1 in gilts and sows was 46.6% (69/148) and 65.1% (241/370), respectively, and that for H3N2 was 78.4% (116/148) and 78.9% (292/370), respectively.

Diagnosis and seroprevalence of porcine respiratory coronavirus disease (돼지호흡기코로나바이러스 감염증의 감별진단과 항체분포 조사)

  • Kim, Eun-Gyeong;Son, Byeong-Kuk;Lee, Jong-Min;Kim, Tho-Kyoung
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.293-298
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    • 2009
  • Porcine respiratory coronavirus (PRCV) is antigenically related to transmissible gastroenteritis virus (TGEV). Differential serological diagnosis between PRCV and TGEV infection is not possible with the classical sero-neutralization test. Infection with PRCV or TGEV induces antibodies which neutralize both viruses to the same titer. However, the enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) can differentiate between PRCV and TGEV infection. This study was carried out to investigate the prevalence of PRCV infection of swine in Gyeongnam province. A total of 391 serum samples from 37 herds in Gyeongnam were examined for antibody to PRCV using blocking ELISA. All serum samples were collected from 130- to 150-day-old pigs between August and December 2006. By ELISA, 182 out of 391 sera tested (46.5%) and 29 out of 37 sample herds (78.4%) were positive against PRCV. Our data suggested that seropositive herds for PRCV are distributed diffusely throughout Gyeongnam. The PCR methods were established to diagnose PRCV spike protein (S) gene. PCR were conducted to identify the PRCV genome against 150 pigs in PRCV antibody positive herds.

Detection of Lymphotropic Herpesviruses by Multiplex Polymerase Chain Reaction

  • Park, Sang-Tae;Kim, Seung-Han;Lee, Dong-Gun;Park, Jung-Hyun;Shin, Wan-Shik;Kim, Tai-Gyu;Paik, Soon-Young;Kim, Chun-Choo
    • Journal of Microbiology
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    • v.39 no.3
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    • pp.226-228
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    • 2001
  • Human lymphotropic herpesvirus is known to be a major pathogen associated with various diseases in bone marrow transplantation (BMT) recipients. A multiplex nested-polymerase chain reaction (PCR) method was developed for the simultaneous detection of human lymphotropic herpesviruses, including Ebstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 variants A and B (HHV6-A, HHV6-B). To demonstrate the usefulness of multiplex PCR for the analysis of clinical samples, peripheral blood mononuclear cells and serum from BMT recipients were analysed. The results skewed that a clear detection could be made between EBV, HCMV and HHV-6. This multiplex PCR assay is an efficient and cost-effective approach to the analysis of large numbers of samples to determine the epidemiological importance of EBV HCMV and HHV-6.

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Nitro oxide in human cytomegalovirus replication and gene expression

  • Lee, Jee-Yeon;Lee, Chan-Hee
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.152-157
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    • 1997
  • Infection of human fibroblast (HF) cells with human cytomegalovirus (HCMV) result in changes in the intracellular level of second messengers. Since nitric oxide (NO) production has been known to be related with other second messengers, it is probable that HCMV infection of HF cells may involve NO. To test this possibility, the amount of NO was measured following ogenous addition of NO generators such as sodium nitroprusside (SNP) or S-nitroso-N-a-cetylpenicillamine (SNAP) immediately after HCMV infection, however, inhibited virus multiplication. Furthermore, immunoblot experiment using monoclonal antibody to HCMV major immediate early (MIE) proteins or CAT assay using pCMVIE/CAT (plasmid containing CAT gene driven by HCMV MIE promoter) revealed that SNP or SNAP blocked the MIE gene expression. SNP was more effective than SNAP in hibiting HCMV multiplication or MIE gene expression. SNP produced more NO than SNAP in inhibiting HCMV multiplication or MIE gene expression. SNP produced more NO than SNAP. Although the mechanism for the inhibition of HCMV multiplication and MIE gene expression by NO is still elusive some correlation with NO-mediated inhibition of HCMV-induced increase in cytosolic free Ca$\^$2+/ concentration ([Ca$\^$2+/]) was observed. The increase of [Ca$\^$2+/] following HCMV infection was inhibited by SNP, and less effectively by SNAP. Raising [Ca$\^$2+/ with bromo-A23187 partially reversed the SNP block of MIE gene expression. Thus, there appear to e some relationships among NO. [Ca$\^$2+/], and HCMV MIE gene expression.

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Investigation of function and regulation of the YB-1 cellular factor in HIV replication

  • Jung, Yu-Mi;Yu, Kyung-Lee;Park, Seong-Hyun;Lee, Seong-Deok;Kim, Min-Jeong;You, Ji-Chang
    • BMB Reports
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    • v.51 no.6
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    • pp.290-295
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    • 2018
  • Y-box binding protein 1 (YB-1) is a member of the cold-shock domain (CSD) protein superfamily. It participates in a wide variety of cellular events, including transcription, RNA splicing, translation, DNA repair, drug resistance, and stress responses. We investigated putative functions of YB-1 in HIV-1 replication. Functional studies using overexpression or knockdown of YB-1 in conjunction with transfection of proviral DNA showed that YB-1 enhances virus production. We found YB-1 regulates HIV-1 production by stimulating viral transcription using HIV-1 LTR sequence U3RU5 with Luciferase assay. We also identified a specific region from amino acids 1 to 324 of YB-1 as necessary for the participation of the protein in the production of virions.

Seroepizootiological survey on bovine leukosis of dairy cattle in Kyunggi province (경기도 지역 유우의 소백혈병 항체 분포 조사)

  • 심항섭;국정희;황영옥;정봉수;김학열;이모란;유성종;강순근;임경애
    • Korean Journal of Veterinary Service
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    • v.21 no.3
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    • pp.255-260
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    • 1998
  • Since bovine leukosis caused considerable economic loss to the dairy industry, seroepidemiologi-cal survey on bovine leukosis was carried out for the dairy herds in Kyunggi province. 1. When compared the results of immunodifussion test with those of enzyme-linked immunosorbent assay(ELISA) for 94 dairy herds sera, the relationship between the immunodifussion test and ELISA were showen high corresponding rate with sensitivity(97.5%) and specificity(92.6%). 2. In immunodiffusion test for bovine leukosis virus (BLV) antibody in 570 dairy cattle from 30 herds, mean positive rate for BLV antibody was 28.2%. The positive rate by districts were 16.5% in central, 35.4% in east, 17.3% in west, 29.1% in south, 31.6% in north, 43.7% in northeast. 3. When the results of serological studies was analyzed by age groups, the number of positive was increased gradually with the advanced in age of herds. The highest positive rate was found in the age over 6 years. 4. Of 30 dairy herds examined, 5 herds(16.7%) have no reactions against BLV antigen while 15 herds (50%) showed the range of 1∼5 positive cattle and 5 herds(16.7%), the rang of over 11 positive cattle.

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Rabies immune status in the stray and companion dogs in Korea

  • Yang, Dong-Kun;Yoon, Soon-Seek;Lee, Kyoung-Ki;Byun, Jae-Won;Bae, You-Chan;Oh, Yoon-I;Song, Jae-Young
    • Korean Journal of Veterinary Research
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    • v.50 no.2
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    • pp.133-137
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    • 2010
  • Rabies virus (family Rhabdoviridae, genus Lyssavirus, RV) is the causative agent of rabies in mammals. We conducted a sero-epidemiological survey for RV using sera from South Korean stray and companion dogs in the present study. A total of 533 canine serum samples were collected between February 2006 and December 2007 and were screened for rabies immunity with a neutralizing peroxidase linked assay. Both companion (49.1%) and stray (60.1%) dogs demonstrated RV seropositivity. Regional RV antibody prevalence was measured in the Jeju (87.5%), Gyeonggi (62%), Gyeongsang (59.1%), Jeonra (42%), Chungcheong (37.9%), and Gangwon (30.4%) provinces. Prevalence increased with age but did not exceed 80% in any age group. Stray and companion dogs had RV antibody prevalence values of 26.7% and 23.7%, respectively. Seroprevalence was significantly associated with age $({\chi}2\;=\;9.46;\;p\;=\;0.024)$ for companion dogs, although this association was not evident in stray dogs. There were no significant differences in age between stray and companion dogs and no gender differences in RV seroprevalence. Our results suggested that a widespread and reinforced vaccination program must be applied to Korean dogs.

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

  • Park Hyo-Sun;Lee Hyo-Sang;Na Ki-Bok;Lee Kwan-Bok;Kang Su-Jung;Moon Sun-Hwa
    • Korean Journal of Veterinary Service
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    • v.29 no.1
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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.

An Effective Method for the Concentration and Detection of Enteroviruses from Water Samples by Combined Cell Culture-Polymerase Chain Reaction (수계 장바이러스의 효과적인 농축과 검출방법의 개발)

  • 장경립;정은영
    • Journal of Life Science
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    • v.10 no.4
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    • pp.368-373
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    • 2000
  • Enteroviruses in the environment pose a public health risk because they can be transmitted via the fecal-oral route through contaminated water, and low numbers are able to initiate an infection in humans. Because the levels of viruses typically found in environmental water and drinking water are low, they must be concentrated from hundreds to thousands of liters of water. Therefore, the main goal of this study was the development of a rapid, simple and efficient procedure to concentrate, isolate and detect enteroviruses from environmental water samples. Viruses were first concentrated by adsorption to 1 MDS cartridge filter and then eluted with approximately 0.5 liter of 1.5% beef extract/0.05M glycin(pH 9.4). In this study, several procedures to concentrate and purify intact viruses from beef extract obtained from the adsorbent filters were tested. Among them, organic floccuration was the best reliable method for reconcentration. sample volume could be reduced to 200∼400 folds and the efficiency of virus recovery through the procedure was over 72%. Finally, the samples were filtered through a membrane disk filter and then analyzed by either the plaque assay or combined cell culture-polymerase chain reaction.

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Construction of tat-and nef-defective HIV-1 and screening of natural extracts with anti-HIV-1 activity

  • Lee, Ann-Hwee;Song, Man-Ki;Suh, Young-Ah;Sung, Young-Chul
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.77-77
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    • 1995
  • Human immunodeficiency virus type 1 (HIV-1) contains several nonstructural genes which are required for the viral replication and disease pathogenesis. Among them, tat and nef genes encode an essential transactivator of HIV-1 LTR and a pluripotent protein which seems to be essential for the in vivo but not in vitro viral replication, respectively. We constructed two tat and n of defective HIV-1 and tested for their ability to replicate in several T cells. The defective viruses did not replicate in CD4$\^$+/ T cells, but rescued in the recombinant Jurkat-tat cell which also contains tat gene. The replication of tat and nef defective HIV-1 which expresses chloramphenicol acetyltransferase(CAT) gene was easily detected by a sensitive CAT assay. No revertant was identified during the passages of the mutant viruses for more than two months in Jurkat-tat cells. tat and n of defective HIV-1 could be used instead of wild type viruse for several purposes such as inhibitor screening and development of attenuated AIDS vaccine.

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