臺灣博碩士論文加值系統

腸病毒腦膜腦炎是最常見的無菌性腦膜腦炎，其預後通常良好且死亡率低；但是腸病毒71型引起之手口足病或疱疹性咽峽炎而併發之腦炎自87年在台灣爆發後，因其侵犯中樞神經易造成神經性肺水腫而有高死亡率。診斷腸病毒71型腦炎主要依照個案的臨床症狀、脊髓液及影像特徵作初步診斷。而確定診斷之工具包括了病毒培養、血清學及逆轉錄聚合酵素鏈鎖反應。這篇主要的目的是檢視以泛腸病毒及腸病毒71型之引子以逆轉錄聚合酵素鏈鎖反應(RT-PCR)在應用在不同檢體中之陽性率性比較。這裡亦回顧了腸病毒71型腦炎之臨床表現及其在神經核磁共振影像之發現。
本研究以90年1月至94年12月因感染腸病毒而引起中樞神經症狀，而進入中國醫藥大學附設醫院兒童加護病房接受治療之兒童為對象，而收集其檢體做分析。每一位均收集四組成對檢體，包括喉頭/肛門拭體、脊髓液及血液，一套以RT-PCR偵測腸病毒基因之陽性與否；另一套則以傳統病毒培養而偵測之。
共有66位符合診斷定義病童，其檢體執行RT-PCR及傳統病毒培養；結果可發現RT-PCR在不同檢體中之陽性率在喉頭(74.2%)及肛門拭體(68.2%)遠高於脊髓液(34.9%)及血液(27.7%)；而在病毒培養亦可發現其陽性率在喉頭(30.3%)及肛門拭體(16.7%)高於脊髓液(1.5%)及血液(3.0%)。而在各不同檢體中，也可以發現RT-PCR之陽性率均明顯高於病毒培養。而以四組檢體做RT-PCR則可鑑別出92%的病童為腸病毒感染，其中65%可以次分型確定為腸病毒71型。
我們結論RT-PCR之陽性率明顯優於病毒培養，其在臨床上是可做為良好鑑別性之診斷工具。

Enteroviral meningoencephalitis is the most prevalent meningoencephalitis, which usually subsequent a well prognosis and low mortility;but enterovirus 71 infections manifested as hand-foot-mouth disease or herpangina with encephalitis that subsequent neurological pulmonary edema and substantial mortality in Taiwan since its breakout in 1998. Tentative diagnosis of enterovirus 71 meningoencephalitis depends on individual clinical symptoms, cerebral spinal fluid (CSF) and neurological images. Final confirmation of virus infection includes virus culture, reverse transcription-polymerase chain reaction (RT-PCR) and serology. The aim of this study was to speculate the application of RT-PCR with universal pan-enteroviral and enteroviral 71(EV71) type- specific primers for direct detections of enterovirus in clinical specimens. Here also review the clinical manifestation and findings of neurological MR imagings in enterovirtus 71 encephalitis.
This project enrolled patients diagnosed of erterovirus central nerve system infection, who admitted to the pediatric intensive care unite of China Medical University Hospital between 2000,Jan to 2005,Dec.Four paired of specimens, including throat/recta swabs, cerebral spinal fluid and blood were collected when admitted; one set were sent for RT-PCR and another were for virus culture.
There were total sixty-six patients who met the diagnostic criteria, with specimens conducted both RT-PCR and virus culture. The results showed the positive rate of RT-PCR were higher in throat swabs (74.2%), rectum swabs (68.2%) than in CSF (34.9%) and blood (27.7%). The same results also showed in virus culture, which consisted of throat swabs (30.3%), rectum swabs (16.7%), CSF (1.5%) and blood (3.0%).Also the positive rate in RT-PCR were demonstrated superior to virus culture in each paired of specimens. Identification of enterovirus infection with RT-PCR was as high as 92% with four specimens, in which 65% were subtyped as enterovirus 71.
We concluded the sensitivity of RT-PCR is superior to viral culture, which works with a well identification in clinical diagnosis of pan-EV and EV71 infection.

中文摘要 ---------------------------------------------------- 1.
英文摘要 -----------------------------------------------------3.
致謝 -------------------------------------------------------- 5.
關鍵字--------------------------------------------------------6.
目錄 ---------------------------------------------------------7.
圖目錄 ------------------------------------------------------10.
表目錄 ------------------------------------------------------12.
第一章 前言
第一節 研究緣起 -----------------------------------------13.
第二節 研究目的-----------------------------------------14.

第二章 文獻探討
第一節 1998腸病毒71型在台灣的爆發---------------------- 16.
第二節 腸病毒71型病理特徵-------------------------------17.
第三節 腸病毒71型病理機轉-------------------------------17.
第四節 腸病毒 71 型腦炎的分期--------------------------- 18.
第五節 腸病毒 71 型腦炎臨床表現--------------------------19.
第五節 聚合酵素鏈鎖反應----------------------------------20.

第三章 研究架構與研究設計
第一節 研究時間及對象------------------------------------21.
第二節 腦幹腦炎之定義----------------------------------- 21.
第三節 檢體採得----------------------------------------- 21.
第四節 核磁共振----------------------------------------- 22.

第四章 研究材料及研究方法
第一節 病毒培養
4-1-1病毒培養之單層細胞株---------------------------23.
4-1-2病毒培養之單層細胞株---------------------------23.
第二節 反轉錄聚合酵素鏈鎖反應
4-2-1反轉錄聚合酵素鏈鎖反應之檢體處置---------------24.
4-2-2反轉錄聚合酵素鏈鎖反應之原理-------------------24.
4-2-3反轉錄聚合酵素鏈鎖反應之步驟-------------------25.
4-2-4反轉錄聚合酵素鏈鎖反應之引子------------------ 25.
4-2-5 3% agarose gel電泳-------------------------- 26.
4-2-6資料收集及統計方法------------------------------26
4-2-7人體試驗委員會----------------------------------26
第五章 研究結果
第一節 腸病毒71型腦炎臨床特徵
5-1-1基本資料---------------------------------------27
5-1-2 腸病毒71型腦炎臨床徵狀------------------------27
第二節 病毒培養及逆轉錄聚合酵素鏈鎖反應之比較
5-2-1 病毒培養及逆轉錄聚合酵素鏈鎖反應之敏感性--------28.
5-2-2 以病毒培養及逆轉錄聚合酵素鏈鎖反應比較診斷
pan-EV及EV71病毒腸病毒71型之敏感性---------- 28.
5-2-3 以病毒培養及逆轉錄聚合酵素鏈鎖反應比較其時效性- 28.
第三節 影像特徵--------------------------------------------29
第六章 討論
第一節 腸病毒中樞神經系統感染症狀----------------------- 30.
第二節 病毒培養及逆轉錄聚合酵素鏈鎖反應之敏感性比較------32.
第三節 神經學影像--------------------------------------- 33.
第七章 結論------------------------------------------------ 34.
參考文獻----------------------------------------------------- 45
作者簡歷及著作------------------------------------------------51

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