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研究生:管宜甄
研究生(外文):Yi-Chen Kuan
論文名稱:應用PCR來檢測兒科呼吸道感染住院病患的病毒性致病因-特別著重在傳統病毒培養無法鑑定出的病毒
論文名稱(外文):APPLICATION OF PCR TO INVESTIGATE THE VIRAL ETIOLOGY OF RESPIRATORY TRACT INFECTION AMONG PEDIATRIC INPATIENTS WITH EMPHASIS ON PATHOGENS NOT IDENTIFIED BY CONVENTIONAL VIRAL CULTURE
指導教授:段國仁段國仁引用關係
指導教授(外文):Kow-Jen Duan
口試委員:段國仁
口試委員(外文):Kow-Jen Duan
口試日期:2015-07-28
學位類別:碩士
校院名稱:大同大學
系所名稱:生物工程學系(所)
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:54
中文關鍵詞:呼吸道感染PCR檢測
外文關鍵詞:RESPIRATORY TRACT INFECTIONPCR INVESTIGATE
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急性病毒性呼吸道感染為兒童最常見的感染症之一,亦是造成嬰幼兒及兒童健康問題的主因。嬰幼童常見的呼吸道感染症狀包括扁桃腺發炎(咽炎)、一般感冒、哮吼、細支氣管炎以及肺炎。除了傳統常見的腺病毒、流行性感冒病毒、副流行性感冒病毒、呼吸道融合病毒這些呼吸道病毒,許多陸續被鑑定出的新型呼吸道病毒諸如:人類肺間質病毒(hMPV)、人類冠狀病毒–NL63(HCoV–NL63)以及人類博卡病毒(HBoV),在2001年起陸續被鑑定出來是造成兒童呼吸道感染的致病原因。如同其他的感染病症,病毒造成的呼吸道感染症其流行病學往往也因地區、國家的不同而不同。我們利用因呼吸道感染症住院且採其鼻咽部檢體做病毒學檢驗的病童,以即時定量聚合酶鏈反應(RT–PCR)來鑑定人類肺間質病毒(hMPV)、人類冠狀病毒–NL63(HCoV–NL63)以及人類博卡病毒(HBoV)這些新鑑定出來的病毒。除了傳統的檢體組織培養方式,針對新鑑定出來的病毒包括鼻病毒(Rhinovirus)、 人類肺間質病毒(hMPV)、人類冠狀病毒–NL63,–OC43,–229E(HCoV–NL63,HCoV–OC43,HCoV–229E)以及人類博卡病毒(HBoV),亦利用即時定量聚合酶鏈反應(RT–PCR)來鑑定。
Acute viral respiratory tract infections (RTIs) are among the most common infections in children and are the major health problems in infants and children. In addition to traditional viral pathogens, including adenoviruses, parainfluenza viruses, influenza viruses, and respiratory syncytial virus (RSV), many respiratory viruses such as human metapneumovirus (hMPV), human coronavirus (HCoV) NL63, coronavirus HKU1, and human bocavirus (HBoV) have been recently (since 2001) identified as causative agents of RTIs in children. A disease entity can be caused by various viruses, and a single virus can also cause various disease entities. As for other infectious diseases,the epidemiological patterns of viral RTIs usually differ between countries and areas.we identified the clinical manifestations of childhood RTIs due to these newly identified respiratory viruses in Taiwan and investigated the comprehensive viral etiology of bronchiolitis and croup in Taiwan. Hospitalized children with respiratory tract infections were randomly selected (about 1 in 10 and a total of 1000 subjects), and nasopharyngeal aspirates were obtained for viral etiologic investigation. The subjects were categorized according to clinical manifestations. For those with croup and bronchiolitis, we attempted to establish a comprehensive viral etiology. In addition to conventional tissue culture and antigen detection, a multiplex polymerase chain reaction method was also conducted for the detection of certain respiratory viruses, including rhinovirus, hMPV, HCoV–229E, HCoV–OC43, HCoV–NL63, and HBoV.
誌謝I
摘要II
ABSTRACTIV
目次VII
表次X
圖次XI
第一章 研究方向及目的1
第二章 背景與目的6
2.1 哮吼(Croup)6
2.2 細支氣管炎(Bronchiolitis)8
2.3 呼吸道感染(Respiratory tract infection,RTIs)9
2.4 兒童呼吸道病毒10
2.5 常見病毒種類14
2.5.1 流行性感冒病毒(Influenza virus)14
2.5.2 副流行性感冒性病毒(Parainfluenza viruses)14
2.5.3 腺病毒(Adenovirus)15
2.5.3.1 腺病毒(Adenovirus)相關的呼吸道感染症候群16
2.5.4 呼吸道融合病毒(Respiratory Syncytial Virus,RSV)17
2.5.5 腸病毒(Enterovirus)19
2.5.6 人類冠狀病毒(Human Coronavirus,HCoV)20
2.5.6.1 人類冠狀病毒–OC4及–229E(Human Coronavirus,HCoV–OC43、–229E)20
2.5.6.2人類冠狀病毒–NL63 Human Coronavirus,HCoV–NL63)21
2.5.7 人類博卡病毒(Human Bocavirus,HBoV)21
2.5.8 人類間質肺炎病毒(Human Metapneumovirus,hMPV)2
2.5.9 鼻病毒(Rhinovirus)22
2.6 近年來相關文獻23
第三章 材料與方法26
3.1 實驗儀器26
3.2 實驗試劑及耗材6
3.3 檢體選擇28
3.4 檢體採集及病毒分離29
3.5 病毒培養29
3.6 細胞病變效應(Cytopathic effects,CPE) 觀察30
3.7 抗體螢光染色(Immunofluorescence assay,IFA)篩檢31
3.8 即時定量聚合酶鏈反應(RT–PCR)31
第四章  實驗結果38
4.1 研究族群背景資料38
4.2 哮吼病患上呼吸道病毒檢驗的分析39
4.3 副流行性感冒病毒佔呼吸道感染的最大比例41
4.4 病毒感染前後之細胞型態差異42
4.5 人類間質肺炎病毒(hMPV)為呼吸道感染的最常見病原43
4.6 呼吸道感染之病毒種類與季節無明顯關聯性8
第五章 討論49
參考文獻51
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