DC 欄位 | 值 | 語言 |
dc.contributor | 蔡向榮 | zh-TW |
dc.contributor | Tsai, Hsiang-Jung | en |
dc.contributor | 臺灣大學:獸醫學研究所 | zh-TW |
dc.contributor.author | 林郁尊 | zh-TW |
dc.contributor.author | Lin, Yu-tsun | en |
dc.creator | 林郁尊 | zh-TW |
dc.creator | Lin, Yu-tsun | en |
dc.date | 2009 | en |
dc.date.accessioned | 2010-05-04T08:26:27Z | - |
dc.date.accessioned | 2018-07-09T06:23:41Z | - |
dc.date.available | 2010-05-04T08:26:27Z | - |
dc.date.available | 2018-07-09T06:23:41Z | - |
dc.date.issued | 2009 | - |
dc.identifier.other | U0001-3007200908480500 | en |
dc.identifier.uri | http://ntur.lib.ntu.edu.tw//handle/246246/178958 | - |
dc.description.abstract | 野鳥一直以來便被認為是A型流感的自然保毒宿主。因此,牠們總是在禽流感的流行病學議題中扮演著相當重要的角色。這個研究嘗試了解 H4N6病毒在台灣野鴨與家鴨及雞隻間的傳播。我們針對 1998年至2007年間分離之10 株台灣野鴨株以及1999年、2006年至2007年間分離之12株台灣家鴨株定序並進行隨後的分析,所有序列資料與基因庫中的H4N6病毒、台灣H6亞型病毒和部分H5亞型病毒之基因序列進行比對分析。結果得到,所有的台灣家鴨株,部分或全部的基因都與台灣野鴨株的基因被分類在親緣關系樹的同一分支中,而家鴨株與野鴨株的 H4 或 N6 亞型病毒的核酸序列最高相似度可達99.9 % (範圍從87.1 %到99.9 %)。這些分析指出台灣野鴨與家鴨間的傳播是可能存在的,因此,當野鴨攜帶病毒飛經台灣時,家鴨將可能暴露在感染的風險下。而在H4亞型病毒的親緣關係樹下,2006年與 2007 年的家鴨分離株並沒有在同一分支中,這顯示 2006 年在家鴨族群中的毒株可能已逐漸消失,無法穩定存在於家鴨族群中。所有的台灣H4N6 分離株在親緣關係樹中都有與其他歐亞地區的分離株分配在同一分支的發現,沒有台灣分離株獨有的分支存在。在基因重組分析中,台灣H4N6分離株的基因重組發生率為4/169 (2.4%),顯示出基因重組在禽類發生的重要性。在血清學調查中,我們發現有小部分的雞血清樣本 (7/8403; 0.08 %) 在H4的 HI 測試下呈低力價陽性反應( 16X ~ 32X),但在2007年雞隻的採樣調查中,並沒有從雞隻分離到任何的H4亞型病毒,這可能表示台灣的極少數雞隻雖可能曾經暴露於H4亞型病毒,但似乎沒有顯著的跨物種傳播在台灣發生。在胺基酸序列分析中,重點比較各基因片段中具有影響力的胺基酸位置,除了A/Wild Duck/Taiwan/147/2001和A/Duck/Taiwan/648/2006這兩株病毒的N6基因發生缺失外,沒有發現其他有義意的突變。然而,本研究藉著codon-based substitution models進行正向選擇位點的分析,找到N6基因上的第285和287個胺基酸位置與NP基因上第105個胺基酸位置可能是因選擇性壓力所造成的正向選擇位點,這些偵測出的特別的胺基酸位點可能有助於了解禽流感病毒在家鴨與野鴨間的演化與選擇性壓力。 | zh-TW |
dc.description.abstract | Wild birds were considered as natural reservoir of influenza A viruses. Hence, they always play important roles in the issue for epidemiology of avian influenza. This study sought to understand the transmission of H4N6 viruses between wild birds and domestic birds in Taiwan. We sequenced total of eight genome segments of ten H4N6 viruses isolated from wild ducks in Taiwan between 1998 and 2007 and twelve H4N6 viruses isolated from domestic ducks in Taiwan in 1999, 2006 and 2007. Sequences mentioned above are analyzed and compared with the sequences of H4N6 viruses, Taiwanese isolates and H5 subtype viruses available in the public database. All of Taiwanese domestic duck isolates, part or all of eight genome sequences are classified in the same sublineage with the sequences of Taiwanese wild duck isolates in phylogenetic trees. The similarity of H4 and N6 nucleotide sequences between wild duck and domestic duck isolates can be as high as 99.9 % (range from 87.1 to 99.9 %). These analyses indicated that the transmission of H4N6 viruses between wild ducks and domestic ducks in Taiwan might exist. Therefore, the domestic duck will be exposed to the risk of influenza virus infection when the wild ducks carrying viruses pass through. In H4 phylogenetic tree, the Taiwanese domestic isolates isolated in the year of 2006 and 2007 did not fall under the same sublineage, which is probably due to the fact that the virus had already disappeared in domestic duck populations in 2006. All Taiwanese H4N6 isolates were found in the sublineages with Eurasian isolates, not form an independent sublineage. In recombination analysis, the rate of recombination of Taiwanese H4N6 isolates was 4/169 (2.4%). In serology survey in Taiwan, we found that some chicken serum samples (7/8403; 0.08 %) have low HI antibody titers ( 16X ~ 32X ) for H4. It indicated that chicken in Taiwan might have been exposed to H4 virus but without significant inter-species transmission. Besides, positive-selection analyses were performed by using codon-based substitution models and found residues 285 and 287 of N6 gene and residue 105 of NP gene are possible positive selection sites. These specific codons detected might be helpful in clarifying the driving force of wild duck and domestic duck influenza virus evolution and in selecting specific targets for selective pressure. | en |
dc.description.tableofcontents | 目錄謝------------------------------------------------------------------------------------------I文摘要-----------------------------------------------------------------------------------------II文摘要--------------------------------------------------------------------------------------IV錄------------------------------------------------------------------------------------------VI目錄-------------------------------------------------------------------------------------------XI目錄-----------------------------------------------------------------------------------------XIV錄------------------------------------------------------------------------------------------XV一章 緒言------------------------------------------------------------------------------------1二章 文獻回顧----------------------------------------------------------------------------3-1家禽流行性感冒病毒特性-----------------------------------------------------------------3-1.1病毒分類及構造----------------------------------------------------------------------3-1.2流行性感冒病毒蛋白質結構及功能--------------------------------------------3 2-1.2.1 血球凝集素--------------------------------------------------------------3 2-1.2.2 神經胺酸酶-----------------------------------------------------------------5 2-1.2.3 核蛋白--------------------------------------------------------------------6 2-1.2.4 基質蛋白---------------------------------------------------------------6 2-1.2.5 非結構蛋白-----------------------------------------------------------------7 2-1.2.6 聚合酶蛋白---------------------------------------------------------------7-1.2.7 PB1-F2蛋白---------------------------------------------------------------8-1.3胺基酸位置與病毒特性的相關性----------------------------------------------8 2-1.3.1 宿主特異性--------------------------------------------------------------8 2-1.3.2 病毒病原性-------------------------------------------------------------9 2-1.3.3 病毒抗藥性-------------------------------------------------------------10-1.4 流行性感冒病毒的複製--------------------------------------------------------11-2 家禽流行性感冒病毒之流行病學-------------------------------------------------12-2.1 病毒傳播途徑及病徵---------------------------------------------------------------12-2.2 A型流行性感冒病毒的自然保毒宿主-----------------------------------------13-2.3 A型流行性感冒感受性之動物-------------------------------------------------142-2.3.1 野鳥--------------------------------------------------------------------152-2.3.2 經濟禽鳥-----------------------------------------------------------------152-2.3.3 鴿子-------------------------------------------------------------------162-2.3.4 豬---------------------------------------------------------------------------172-2.3.5 貓-------------------------------------------------------------------------172-2.3.6 狗-------------------------------------------------------------------------182-2.3.7 海豹、鯨魚------------------------------------------------------------182-2.3.8 貂科動物--------------------------------------------------------------192-2.3.9 老鼠----------------------------------------------------------------------19-2.4 家禽流行性感冒病毒H4亞型之流行病學-------------------------------------19-3 流行性感冒病毒抗原變異性與基因分析之應用-------------------------------22-3.1 流行性感冒病毒抗原變異性-----------------------------------------------------22-3.1.1 抗原漂移(Antigen drift) -------------------------------------------------22-3.1.2 抗原轉換(Antigen shift; reassortment) ----------------------------------22-3.1.3 基因重組(Recombination)----------------------------------------------22-3.2 流行性感冒病毒之基因分析---------------------------------------------------23-3.2.1 親緣性分析 (Phylogenetic analysis) ----------------------------------23-3.2.2 胺基酸序列分析---------------------------------------------------------24-3.3 流行性感冒病毒之演化分析---------------------------------------------------242-3.3.1 演化學---------------------------------------------------------------------242-3.3.2 中性理論----------------------------------------------------------------252-3.3.3 天擇模式--------------------------------------------------------------252-3.3.4 演化分析----------------------------------------------------------------26 2-3.3.4.1非同義鹼基替代及同義鹼基替代----------------------------26 2-3.3.4.2 Tajima’s D test--------------------------------------------------27 2-3.3.4.3 Heterogeneity test----------------------------------------------27 2-3.3.4.4 Codon-based substitution models----------------------------27三章 材料與方法---------------------------------------------------------------------------29-1 H4亞型血清學調查---------------------------------------------------------------------29-1.1 樣本來源--------------------------------------------------------------------------29-1.2 血清樣本抗體檢測-----------------------------------------------------------------29 3-1.2.1 製備紅血球溶液-----------------------------------------------------------29 3-1.2.2 H4亞型血球凝集抑制試驗----------------------------------------------29-2. 家禽流行性感冒病毒之基因分析------------------------------------------------30-2.1採樣及運輸與儲存---------------------------------------------------------30-2.2 RNA病毒之萃取---------------------------------------------------------------30-2.3反轉錄反應----------------------------------------------------------------------31-2.4 聚合酶鏈反應------------------------------------------------------------------31-2.5 檢體的處理-------------------------------------------------------------------32-2.6 病毒增殖--------------------------------------------------------------32-2.7 病毒------------------------------------------------------------------------33-2.8 增幅PB2、PB1、PA、HA、NP、NA、MP、NS八段基因全長------------33-2.9 瓊膠醣凝膠電泳----------------------------------------------------34-2.10聚合酶鏈反應產物之純化--------------------------------------34-2.11核酸序列之定序---------------------------------------------------35-2.12病毒核酸序列之分析-------------------------------------------------35-2.13 胺基酸位置定序-----------------------------------------------36-2.14 偵測正向選擇位點------------------------------------------------- 37四章 結果---------------------------------------------------------------------------38-1 H4亞型流行病學調查---------------------------------------------------------------38-1.1 H4亞型病毒分離-----------------------------------------------------------38-1.2 H4亞型血清學調查------------------------------------------------------38-2 以反轉錄聚合酶鏈反應增幅八段基因全長之結果----------------------------38-3 家禽流行性感冒病毒核酸序列之分析-------------------------------------------40-3.1 H4N6亞型核酸序列及胺基酸序列之相似度-----------------------------40-3.1.1 H4基因-------------------------------------------------------------------40-3.1.2 N6基因-------------------------------------------------------------------40-3.1.3 M基因---------------------------------------------------------------------41-3.1.4 NP基因---------------------------------------------------------------------41-3.1.5 NS基因-------------------------------------------------------------------41-3.1.6 PA基因-----------------------------------------------------------------42-3.1.7 PB1基因-----------------------------------------------------------------42-3.1.8 PB2基因-------------------------------------------------------------------43-3.1.9 DK5568-06和WD904-06-----------------------------------------43-3.2 H4N6亞型親原樹狀圖-------------------------------------------------------44-3.2.1 H4基因-------------------------------------------------------------44-3.2.2 N6基因------------------------------------------------------------------44-3.2.3 M基因-------------------------------------------------------------------45-3.2.4 NP基因------------------------------------------------------------------45-3.2.5 NS基因--------------------------------------------------------------------45-3.2.6 PA基因---------------------------------------------------------------------46-3.2.7 PB1基因---------------------------------------------------------------46-3.2.8 PB2基因------------------------------------------------------------------47-3.3基因重組分析-------------------------------------------------------------------47-3.4 H4N6亞型胺基酸位置之分析--------------------------------------------------48-3.5偵測正向選擇位點-------------------------------------------------------------49五章 討論-------------------------------------------------------------------------------50六章 參考文獻--------------------------------------------------------------------------91錄---------------------------------------------------------------------------------------------102 | en |
dc.format | application/pdf | en |
dc.format.extent | 4810636 bytes | - |
dc.format.mimetype | application/pdf | - |
dc.language | zh-TW | en |
dc.language.iso | en_US | - |
dc.subject | H4N6亞型病毒 | zh-TW |
dc.subject | 跨物種傳播 | zh-TW |
dc.subject | 親緣關係分析 | zh-TW |
dc.subject | 基因重組 | zh-TW |
dc.subject | 血清學調查 | zh-TW |
dc.subject | H4N6 subtype | en |
dc.subject | inter-species transmission | en |
dc.subject | phylogenetic analysis | en |
dc.subject | recombination | en |
dc.subject | serological survey | en |
dc.title | 台灣家鴨及野鴨H4N6亞型流行性感冒病毒流行病學分析 | zh-TW |
dc.title | Epidemiological analysis of H4N6 viruses isolated from wild ducks and domestic ducks in Taiwan | en |
dc.type | thesis | en |
dc.identifier.uri.fulltext | http://ntur.lib.ntu.edu.tw/bitstream/246246/178958/1/ntu-98-R96629039-1.pdf | - |
item.fulltext | with fulltext | - |
item.languageiso639-1 | en_US | - |
item.openairecristype | http://purl.org/coar/resource_type/c_46ec | - |
item.cerifentitytype | Publications | - |
item.openairetype | thesis | - |
item.grantfulltext | open | - |
顯示於: | 獸醫學系
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