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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 高全良(Chuan-Liang Kao) | |
| dc.contributor.author | Jo-Shan Wang | en |
| dc.contributor.author | 王若珊 | zh_TW |
| dc.date.accessioned | 2021-06-15T01:13:11Z | - |
| dc.date.available | 2014-09-15 | |
| dc.date.copyright | 2009-09-15 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-07-29 | |
| dc.identifier.citation | 1.Allander, T., et al., Cloning of a human parvovirus by molecular screening of respiratory tract samples. Proc Natl Acad Sci U S A, 2005. 102(36): p. 12891-6.
2.Durham, P.J., et al., Epidemiological studies of parvovirus infections in calves on endemically infected properties. Res Vet Sci, 1985. 38(2): p. 234-40. 3.Durham, P.J., A. Lax, and R.H. Johnson, Pathological and virological studies of experimental parvoviral enteritis in calves. Res Vet Sci, 1985. 38(2): p. 209-19. 4.Fields, B.N., D.M. Knipe, and P.M. Howley, Fields' virology. 5th ed. 2007, Philadelphia: Wolters Kluwer Health/Lippincott Williams & Wilkins. 2 v. (xix, 3091, 86 p.). 5.Schildgen, O., et al., Human bocavirus: passenger or pathogen in acute respiratory tract infections? Clin Microbiol Rev, 2008. 21(2): p. 291-304, table of contents. 6.Carmichael, L.E., D.H. Schlafer, and A. Hashimoto, Pathogenicity of minute virus of canines (MVC) for the canine fetus. Cornell Vet, 1991. 81(2): p. 151-71. 7.Manteufel, J. and U. Truyen, Animal Bocaviruses: A Brief Review. Intervirology, 2008. 51(5): p. 328-334. 8.Allander, T., et al., Human bocavirus and acute wheezing in children. Clin Infect Dis, 2007. 44(7): p. 904-10. 9.Arnold, J.C., et al., Human bocavirus: prevalence and clinical spectrum at a children's hospital. Clin Infect Dis, 2006. 43(3): p. 283-8. 10.Chung, J.Y., et al., Bocavirus infection in hospitalized children, South Korea. Emerg Infect Dis, 2006. 12(8): p. 1254-6. 11.Naghipour, M., et al., Human bocavirus in Iranian children with acute respiratory infections. J Med Virol, 2007. 79(5): p. 539-43. 12.Qu, X.W., et al., Human bocavirus infection, People's Republic of China. Emerg Infect Dis, 2007. 13(1): p. 165-8. 13.Sloots, T.P., et al., Evidence of human coronavirus HKU1 and human bocavirus in Australian children. J Clin Virol, 2006. 35(1): p. 99-102. 14.Vicente, D., et al., Human bocavirus, a respiratory and enteric virus. Emerg Infect Dis, 2007. 13(4): p. 636-7. 15.Neske, F., et al., Real-time PCR for diagnosis of human bocavirus infections and phylogenetic analysis. J Clin Microbiol, 2007. 45(7): p. 2116-22. 16.Chieochansin, T., et al., Human bocavirus infection in children with acute gastroenteritis and healthy controls. Jpn J Infect Dis, 2008. 61(6): p. 479-81. 17.Campe, H., C. Hartberger, and A. Sing, Role of Human Bocavirus infections in outbreaks of gastroenteritis. J Clin Virol, 2008. 43(3): p. 340-2. 18.Ziegler, S., et al., No gastroenteric Bocavirus in high risk patients stool samples, in J Clin Virol. 2008. p. 349-50. 19.Lee, J.I., et al., Detection of human bocavirus in children hospitalized because of acute gastroenteritis. J Infect Dis, 2007. 196(7): p. 994-7. 20.Kapoor, A., et al., A Newly Identified Bocavirus Species in Human Stool. J Infect Dis, 2009. 199(2): p. 196-200. 21.Arthur, J.L., et al., A novel bocavirus associated with acute gastroenteritis in Australian children. PLoS Pathog, 2009. 5(4): p. e1000391. 22.Chieochansin, T., et al., Complete coding sequences and phylogenetic analysis of Human Bocavirus (HBoV). Virus Res, 2007. 129(1): p. 54-7. 23.Bastien, N., et al., Detection of human bocavirus in Canadian children in a 1-year study. J Clin Microbiol, 2007. 45(2): p. 610-3. 24.Lau, S.K., et al., Clinical and molecular epidemiology of human bocavirus in respiratory and fecal samples from children in Hong Kong. J Infect Dis, 2007. 196(7): p. 986-93. 25.Chieochansin, T., et al., Human bocavirus (HBoV) in Thailand: clinical manifestations in a hospitalized pediatric patient and molecular virus characterization. J Infect, 2008. 56(2): p. 137-42. 26.Manning, A., et al., Comparison of tissue distribution, persistence, and molecular epidemiology of parvovirus B19 and novel human parvoviruses PARV4 and human bocavirus. J Infect Dis, 2007. 195(9): p. 1345-52. 27.Garbino, J., et al., Respiratory viruses in HIV-infected patients with suspected respiratory opportunistic infection. AIDS, 2008. 22(6): p. 701-5. 28.Kleines, M., et al., High prevalence of human bocavirus detected in young children with severe acute lower respiratory tract disease by use of a standard PCR protocol and a novel real-time PCR protocol. J Clin Microbiol, 2007. 45(3): p. 1032-4. 29.Lu, X., et al., Real-time PCR assays for detection of bocavirus in human specimens. J Clin Microbiol, 2006. 44(9): p. 3231-5. 30.Schenk, T., et al., Human bocavirus DNA detected by quantitative real-time PCR in two children hospitalized for lower respiratory tract infection. Eur J Clin Microbiol Infect Dis, 2007. 26(2): p. 147-9. 31.Fry, A.M., et al., Human bocavirus: a novel parvovirus epidemiologically associated with pneumonia requiring hospitalization in Thailand. J Infect Dis, 2007. 195(7): p. 1038-45. 32.Kantola, K., et al., Serodiagnosis of human bocavirus infection. Clin Infect Dis, 2008. 46(4): p. 540-6. 33.Brieu, N., et al., Electron microscopy observation of human bocavirus (HBoV) in nasopharyngeal samples from HBoV-infected children. J Clin Microbiol, 2007. 45(10): p. 3419-20. 34.Lin, F., et al., ELISAs using human bocavirus VP2 virus-like particles for detection of antibodies against HBoV. J Virol Methods, 2008. 149(1): p. 110-7. 35.Endo, R., et al., Seroepidemiology of human bocavirus in Hokkaido prefecture, Japan. J Clin Microbiol, 2007. 45(10): p. 3218-23. 36.Qu, X.W., et al., Phospholipase A2-like activity of human bocavirus VP1 unique region. Biochem Biophys Res Commun, 2008. 365(1): p. 158-63. 37.Bohmer, A., et al., Novel application for isothermal nucleic acid sequence-based amplification (NASBA). J Virol Methods, 2009. 158(1-2): p. 199-201. 38.Dijkman, R., et al., Human bocavirus can be cultured in differentiated human airway epithelial cells. J Virol, 2009. 39.Lin, J.H., et al., Clinical and genetic analysis of Human Bocavirus in children with lower respiratory tract infection in Taiwan. J Clin Virol, 2009. 40.Lindner, J., et al., CD4(+) T Helper Cell Responses against Human Bocavirus Viral Protein 2 Viruslike Particles in Healthy Adults. J Infect Dis, 2008. 198(11): p. 1677-84. 41.Lindner, J., et al., Humoral immune response against human bocavirus VP2 virus-like particles. Viral Immunol, 2008. 21(4): p. 443-9. 42.Kahn, J.S., et al., Seroepidemiology of human bocavirus defined using recombinant virus-like particles. J Infect Dis, 2008. 198(1): p. 41-50. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/42402 | - |
| dc.description.abstract | Bocavirus在2005年9月首次由瑞典Karolinska大學醫院的Tobias Allander團隊從有呼吸道不適症狀的孩童及嬰幼兒所發現，分析其基因發現與Bovine Parvovirus及Canine Minute virus分別有42％和43％的相似度，因此命名為Human Bocavirus（HBoV，人類玻卡病毒）[1]。人類玻卡病毒被分類屬於Parvoviridae family（細小病毒科）, Parvovirus subfamily, Bocavirus genus，細小病毒科為DNA病毒當中最小的，特性包含病毒顆粒很小，大約20~25nm，且不具有套膜，其核酸為單股DNA。由基因的分析可知人類玻卡病毒有三個open reading frame，分別轉譯出NS1、NP1、VP1/2蛋白質。HBoV目前在許多國家的呼吸道疾病病人中都有被偵測到，陽性率約為1.5%~19%。先前的研究指出孩童及嬰幼兒感染HBoV的機率比較高。此外，有部份研究調查糞便檢體發現人類玻卡病毒與腸胃炎有些許相關性。利用baculovirus系統表現VP2蛋白質來調查抗體盛行率，小孩的感染率大約30~40%，而成人最高可達95%。
為瞭解台灣地區是否亦有玻卡病毒之存在，因此進行了相關研究。研究設計為針對台灣北部地區有呼吸道感染之臨床檢體進行收集。收集時間為2008年11月到2009年5月，檢測則利用核酸檢測技術。在檢體直接檢測方面，發現北台灣確實有HBoV之存在，總共檢測402件，有31件（7.71%）為陽性，當中最常見的診斷為bronchiolitis，佔了41.94%。從不同月份進行比較可看出從2008年11月份開始採檢，有隨著月份逐漸減少的情形，年齡從0.4~8.5歲皆有。而HBoV感染病患也具有常見的呼吸道症狀如發燒(48.39%)、咳嗽(93.59%)、流鼻水(80.65%)。將陽性檢體萃取之DNA進行NS1、NP1、partial VP region之核酸分析比對，NS1、NP1序列變異很小，而從partial VP種系圖則顯示台灣地區至少有兩種基因型存在，且與其他台灣地區研究中之分型得到一樣的結果。另外使用SYBR Green Real-time PCR定量檢體中的病毒量，臨床檢體的病毒量最低10^2.98到最高10^6.19 copies/μl（平均為10^4.52copies/μl）。由於目前還沒有台灣地區的相關血清流行病學調查，因此也建立利用baculovirus表現系統表現HBoV之VP2蛋白質，以期未來能應用於偵測台灣地區各年齡層的抗體陽性率，以瞭解台灣地區HBoV流行的狀況。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-15T01:13:11Z (GMT). No. of bitstreams: 1 ntu-98-R96424019-1.pdf: 1375175 bytes, checksum: 9842e29aac31599ce5cd4b79fe0fed77 (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 目錄
口試委員審定書....................................i 誌謝..............................................ii 中文摘要..........................................iii 英文摘要..........................................v 第一章 緒論.......................................1 1.1 HBoV的發現..............................1 1.2 MVC.....................................2 1.3 BPV.....................................3 1.4 HBoV的基因體............................5 1.5 HBoV呼吸道感染的關係....................5 1.6 HBoV與腸胃炎............................6 1.7 HBoV的基因變異性........................6 1.8 HBoV與免疫功能不全病患..................6 1.9 HBoV viral load.........................7 1.10 電顯下的HBoV顆粒........................7 1.11 HBoV與其他病毒的共同感染................8 1.12 HBoV之血清盛行率........................8 1.13 HBoV VP1u之phospholipase活性............9 1.14 HBoV顆粒中核酸的特性....................9 1.15 HBoV的培養與mRNA........................9 1.16 台灣地區的HBoV..........................10 1.17 研究目的....................................10 第二章 材料與方法 ................................11 I. 實驗材料......................................11 II. 實驗方法.....................................12 2.1 檢體採集.....................................12 2.2 運用PCR方法偵測檢體中的人類玻卡病毒..........12 2.3 細胞、病毒培養...............................13 2.4 核酸定序.....................................14 2.5 建構HBoV已知全長之質體.......................15 2.6 SYBR Green real time PCR.....................18 2.7 Nest PCR偵測HBoV2............................18 2.8 Baculovirus expression system................19 第三章 結果 3.1 臨床檢體進行HBoV核酸之偵測...................24 3.2 建構含HBoV已知全長基因之質體.................24 3.3 建立SYBR Green Real-Time PCR以定量檢體中病毒核酸 量...........................................25 3.4 以不同細胞嚐試培養HBoV可能性.................25 3.5 分析北台灣地區HBoV的基因序列.................26 3.6 喉頭拭子檢體中沒有偵測到HBoV2之存在..........26 3.7 利用昆蟲細胞表現HBoV病毒顆粒表面蛋白質VP2....27 第四章 討論.......................................28 圖表附錄..........................................32 參考文獻 .........................................48 圖目錄 圖1. 偵測HBoV核酸之敏感性分析.....................32 圖2. HBoV各月份盛行率 ............................33 圖3. HBoV陽性檢體之年齡分佈 ......................34 圖4. 建構HBoV全長clone設計........................35 圖5. 以限制酶切割確認HBoV全長質體之建構...........36 圖6. HBoV SYBR Green Real-time PCR之標準曲線......37 圖7. HBoV陽性檢體viral load之分析 ................38 圖8. HBoV序列與HBoV2之種系圖分析..................39 圖9.HBoV序列之種系圖分析..........................40 圖10.確認Bacmid進行transpose結果 .................41 圖11.測試桿狀病毒HBoV VP2表現情形.................42 表目錄 表1. HBoV陽性檢體臨床診斷統計.....................43 表2. 以不同細胞測試HBoV培養 ......................44 表3. HBoV NS1基因之核酸及胺基酸相似度分析.........45 表4. HBoV NP1基因之核酸及胺基酸相似度分析.........46 表5. HBoV partial VP基因之核酸及胺基酸相似度分析..47 | |
| dc.language.iso | zh-TW | |
| dc.title | 偵測及分析北台灣地區人類玻卡病毒及建立其VP2蛋白質之桿狀病毒表現 | zh_TW |
| dc.title | Detection and Genetic Analysis of Human Bocavirus (HBoV) in Northern Taiwan and Establishment of A Baculovirus Expression System of HBoV VP2 Protein | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李君男,張淑媛 | |
| dc.subject.keyword | 人類玻卡病毒,桿狀病毒表現,種系分析, | zh_TW |
| dc.subject.keyword | human bocavirus,baculovirus expression,phylogenic analysis, | en |
| dc.relation.page | 50 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2009-07-29 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 醫學檢驗暨生物技術學研究所 | zh_TW |
| 顯示於系所單位： | 醫學檢驗暨生物技術學系 | |
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