• Title/Summary/Keyword: viral DNA

Search Result 443, Processing Time 0.024 seconds

Rapid detection of deformed wing virus in honeybee using ultra-rapid qPCR and a DNA-chip

  • Kim, Jung-Min;Lim, Su-Jin;Kim, SoMin;Kim, MoonJung;Kim, ByoungHee;Tai, Truong A;Kim, Seonmi;Yoon, ByoungSu
    • Journal of Veterinary Science
    • /
    • v.21 no.1
    • /
    • pp.4.1-4.9
    • /
    • 2020
  • Fast and accurate detection of viral RNA pathogens is important in apiculture. A polymerase chain reaction (PCR)-based detection method has been developed, which is simple, specific, and sensitive. In this study, we rapidly (in 1 min) synthesized cDNA from the RNA of deformed wing virus (DWV)-infected bees (Apis mellifera), and then, within 10 min, amplified the target cDNA by ultra-rapid qPCR. The PCR products were hybridized to a DNA-chip for confirmation of target gene specificity. The results of this study suggest that our method might be a useful tool for detecting DWV, as well as for the diagnosis of RNA virus-mediated diseases on-site.

Sequence Variations of Hepatitis B Virus Promotor Regions in Vertically Transmitted Mother-child Pairs (수직 감염된 B형 간염 바이러스 Promoter 유전자의 변이 분석)

  • Lee, Choong-Won;Han, Young-Na;Lee, Jung-Hwa;Lee, Kwang-Chul;Ha, Young-Mee
    • Pediatric Gastroenterology, Hepatology & Nutrition
    • /
    • v.5 no.1
    • /
    • pp.39-50
    • /
    • 2002
  • Hepatitis B viral infection which affect about 10% of Korean population manifests asymptomatic carrier, chronic hepatitis and liver cirrhosis and even associates with hepatocellular carcinoma. Clinical manifestations induced by hepatitis B virus vary depending on the degree of immune response by cytotoxic T cells against viral epitope-presenting liver cells. Since hepatitis B virus presents high rate of mutaton that might change the presented epitope and eventually alter immune response, viral mutations, especially in promoters and enhancers, have an important implication in hepatic inflammation and viral replication. To identify mutations related to the hepatic inflammation, we investigated sequence variations of hepatitis B viral promotor regions in the presence or absence of symptoms in hepatitis B carriers. For this, sera from persistently hepatitis B virus-infected mother-child pairs were collected. After PCR amplifiation of all hepatitis B viral promoters (C promoter, S1 promoter, S2/S promoter, X promoter) using serum DNA from each pair, viral promotors were sequenced by automatic sequencer and then sequence data were analyzed by ClustalW. In most cases, the dominant type of maternal virus was transmitted to the child. However, in some children, some new host specific viral variants could be observed in Cp, S1p and S2/Sp. The mutations in C promoter did not seem to be vertically transmitted but arose in new host independently after the wild type had been transmitted. Enhancer I containing X promoter revealed high host specific variations as has been reported before. Two S promoters, S1p and S2/Sp, have shown some point mutations in children, but no deletion mutations were detected as in chronic hepatitis patients in whom deletion mutations are frequently found. In conclusion, the children with the vertically transmitted hepatitis B virus mostly retain the dominant type virus that had been transmitted. However, host specific variants tended to accumulate over time, possibly as clinical symptoms develop.

  • PDF

Researches of Epigenetic Epidemiology for Infections and Radiation as Carcinogen

  • Bae, Jong-Myon
    • Journal of Preventive Medicine and Public Health
    • /
    • v.51 no.4
    • /
    • pp.169-172
    • /
    • 2018
  • In recent years, a number of studies have been reported on the various types of cancer arising from epigenetic alterations, including reports that these epigenetic alterations occur as a result of radiation exposure or infection. Thyroid cancer and breast cancer, in particular, have high cancer burden, and it has been confirmed that radiation exposure or onco-viral infection are linked to increased risk of development of these two types of cancer, respectively. Thus, the environment-epigenetic alteration-cancer occurrence (EEC) hypothesis has been suggested. This paper reviews the trends in research supporting this hypothesis for radiation exposure and onco-viral infection. If more evidences accumulate for the EEC hypothesis from future research, those findings may greatly aid in the prevention, early diagnosis, treatment, and prognosis of the thyroid cancer and breast cancer.

Characterization and RT-PCR Detection of Turnip Mosaic Virus Isolated from Chinese Cabbage in Korea (배추에서 분리한 순무 모자이크 바이러스의 특성 및 역전사 중합효소 연쇄반응법(RT-PCR)을 이용한 검정)

  • 박원목;최설란;김수중;최승국;류기현
    • Korean Journal Plant Pathology
    • /
    • v.14 no.3
    • /
    • pp.223-228
    • /
    • 1998
  • Turnip mosaic virus)TuMV-Ca) was isolated from a Chinese cabbage showing severe mosaic and black necrotic spots symptoms in Korea. The virus was identified as a strain of TuMV by its host range test, particle morphology, serology, double stranded RNA analysis. For detection of the virus, reverse transcription and polymerase chain reaction(RT-PCR) was performed with a set of 18-mer TuMV-specific primers to amplify a 876 bp DNA fragment The virus was rapidly detected from total nucleic acids of virus infected tissues as well as native viral RNA of purified virion particles by RT-PCR. Detection limit of the viral RNA by RT-PCR was 10 fg.

  • PDF

The serodiagnosis of a lymphocystis disease virus infection using an antibody raised against a recombinant major capsid protein

  • Seo, Ja-Young;Kang, Bong-Jo;Oh, Hyoung-Jong;Lee, Jae-Il;Kim, Tae-Jung
    • Journal of fish pathology
    • /
    • v.21 no.3
    • /
    • pp.175-180
    • /
    • 2008
  • Lymphocystis is a viral disease of fish primarily in marine and brackishwaters. Here we report the cloning, expression, and the serological applications of the lymphocystis disease virus (LCDV) major capsid protein (MCP). The MCP gene was amplified by PCR from the genomic DNA of LCDV isolated from Schlegel's black rockfish, Sebastes schlegeli, and expressed in E. coli. Mouse antisera raised against the purified recombinant MCP (rMCP) reacted with the viral MCP in an immunofluorescence assay, indicating that this rMCP would be useful for serological studies of field samples.

A Method of Preparing Recombinant Fusion Antigen from Rotavirus and Norovirus

  • Oh, Ho-Kyung;Huh, Chul-Sung;Baek, Young-Jin;Yoo, Dong-Wan
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
    • /
    • 2004.05a
    • /
    • pp.353-356
    • /
    • 2004
  • Rotavirus and Norovirus are major causative agents of acute diarrhea and gastroenteritis. In our study, Each viral RNA was isolated from the feces of patients for viral diarrhea in Korea, respectively. And cDNA library were constructed using RT-PCR. Also, cDNAs encoding VP8 derived from Rotavirus and Capsid protein derived from norovirus were subesequently cloned and expressed in Echerichia coli as a fusion antigen. Molecular weight of fusion antigen was approximately 60kDa. Also, substantial overexpression was accomplished. We yielded egg yolk lgY which is potentially useful in controlling of Rotavirus and Norovirus which are one of the most prevalent pathogenic viruses.

  • PDF

Analysis of Human and Dengue Viral Proteins Interaction Network for Understanding Viral Pathogenesis (감염경로 탐색을 위한 사람 및 뎅기 바이러스 단백질 상호작용 네트워크 분석)

  • Lee, Jihoo;Kim, Hak Yong
    • Proceedings of the Korea Contents Association Conference
    • /
    • 2016.05a
    • /
    • pp.189-190
    • /
    • 2016
  • 바이러스는 RNA나 DNA의 유전물질과 그것을 둘러싸고 있는 최소한의 단백질들만으로 구성되어 있기 때문에 바이러스가 증식하기 위해서는 숙주세포에 침투하여 전적으로 숙주의 복제 기구를 이용해야만 한다. 하지만 아직까지 뎅기 바이러스의 감염 및 복제 기전은 명확하게 밝혀지지 않았다. 이에 본 연구에서는 바이러스의 감염 및 복제기전에 대한 유용한 정보를 도출하기 위하여 사람 단백질과 뎅기 바이러스 단백질의 상호작용(Hu-DV PPI) 네트워크를 분석하였다. 우선 문헌조사를 통하여 실험적으로 검증된 뎅기 바이러스 단백질(9개)과 상호작용하는 사람 단백질(149개)을 추출하였으며, 이 정보를 이용하여 사람-뎅기 바이러스 단백질 상호작용 네트워크를 구축하였다. 이 네트워크를 기반으로 바이러스 감염 전/후의 네트워크 구조 및 특성을 분석하였으며, 이 정보를 바탕으로 감염경로를 탐색하였다.

  • PDF

Molecular Biological Characterization of Recombinant Baculovirus with an Expanded Host Range (숙주범위가 넓어진 유전자 재조합 핵다각체병 바이러스의 분자생물학적 특성)

  • 김우진;우수동
    • Journal of Sericultural and Entomological Science
    • /
    • v.38 no.1
    • /
    • pp.42-47
    • /
    • 1996
  • To investigate the host range determining factors of nuclear polyhedrois virus (NPV), Autographa california NPV and Bombyx mori NPV were coinfected into the two different cell lines, BmN-4 and Sf-9. The recombinant baculoviruses, RecS-A6 and RecB-727 which have an expanded host range, were isolated from Sf-9 and BmN-4 cell lines, respectively. The molecular biological characteristics of the recombinant baculoviruses were investigated. The pathogenicity of RecB-727 was similar to that of wild type BmNPV, while the pathogenicity of RecS-A6 was relatively lower than that of wild type BmNPV. The restriction enzyme digestion patterns of parental viruses and recombinant viruses showed that the recombinant virus has an expanded host range by genetic recombination. Southern blot analysis revealed that the p10 gene of RecB-727 was derived from AcNPV genomic DNA, while RecS-A6 has p10 gene of BmNPV in a viral genome. To investigate the host range expansion mechanism of recombinant baculovirus, HindIII-SacI 0.6 kb DNA fragments of RecS-A6 and RecB-727 were cloned and sequenced. The results showed that of wild type BmNPV helicase gene, suggesting that the expanded host range of recombinant baculoviruses was due to the insertion of BmNPV helicase gene into AcNPV viral genome.

  • PDF

Hepatic Steatosis: Prevalence and Host/Viral Risk Factors in Iranian Patients with Chronic Hepatitis B Infection

  • Poortahmasebi, Vahdat;Alavian, Seyed Moayed;Keyvani, Hossein;Norouzi, Mehdi;Mahmoodi, Mahmood;Jazayeri, Seyed Mohammad
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.9
    • /
    • pp.3879-3884
    • /
    • 2014
  • Background: In chronic hepatitis B (CHB), the presence of hepatic steatosis (HS) seems to be associated with known host and viral factors which may influence the long-term prognosis of chronic hepatitis B (CHB), probably leading to cirrhosis and hepatocellular carcinoma (HCC). Different from chronic hepatitis C (CHC), factors associated with HS in CHB are not clearly explored. Materials and Methods: 160 CHB patients were divided into two groups depending on the results of liver biopsy. Group I consisted of 71 patients with confirmed steatosis. Group II comprised 89 patients without steatosis. The groups were compared in terms of basal characteristics, body mass index (BMI), liver enzymes (ALT, AST, ALP), serum fasting blood sugar (FBS) and lipids, hepatitis B e antigen (HBeAg), viral load, and histological findings. Results: In terms of host factors, male gender, older age, BMI, high serum FBS and lipid levels were associated with HS. On the other hand, ALT levels, the HAI scores of necroinflammation and stage of fibrosis did not associate with HS. On multivariate analysis, parameters of sex, BMI, cholesterol and FBS levels were independently associated with HS. Regarding viral factors, HBeAg negativity was significantly associated with HS (81.7%, p value 0.006), but not HBV DNA level (p value 0.520). Conclusions: HS in CHB appears to be unrelated to the status of HBV replication. However, fibrosis progression in CHB is related to variable host factors. HS may be enhanced through these factors in HBV chronic patients.

Transcriptional Activator Elements for Curtovirus C1 Expression Reside in the 3' Coding Region of ORF C1

  • Hur, Jingyung;Buckley, Kenneth J.;Lee, Sukchan;Davis, Keith R.
    • Molecules and Cells
    • /
    • v.23 no.1
    • /
    • pp.80-87
    • /
    • 2007
  • Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV), members of curtoviruses, encode seven open reading frames (ORFs) within a ~3 kb genome. One of these viral ORFs, C1, is known to play an important role in the early stage of viral infection in plants during initiation of viral DNA replication. We used promoter:: reporter (${\beta}$-glucuronidase) gene fusions in transgenic Arabidopsis to identify the putative promoter region of BCTV ORF C1. Unlike other geminiviruses, the intergenic region of BCTV was not sufficient to promote C1 expression in transgenic plants. When sequences extending into the coding region of C1 were tested, strong expression of the reporter protein was observed in vascular tissues of transgenic plants. This expression was not dependent on the presence of the intergenic regions or proximal 5' portions of the C1 coding region. Transgenic plants expressing a reporter gene under control of the putative complete C1 promoter were inoculated with virus to determine if any viral transcript affected C1 expression. Virus inoculated plants did not show any altered pattern or change in of reporter gene expression level. These results suggest that (1) important transcriptional activator elements for C1 expression reside in the 3' portion of C1 coding area itself, (2) C1 protein does not auto-regulate its own expression and (3) C1 expression of two curtoviruses is controlled differently compared to other geminiviruses.