- 學位論文
CODEHOP分子技術用於腸病毒檢驗之評估
The application of CODEHOP molecular technology in the clinical diagnosis of Enterovirus
摘要
全世界每年有超過五十億個腸病毒感染病例發生，導致患者出現不明熱（undifferentiated febrile illness）、紅疹、腸胃道疾病、無菌性腦膜炎、肢體麻痺、或嚴重的新生兒敗血症。1998年腸病毒在全台灣造成重大疫情，導致七十八位因腦幹受到侵襲進而引發肺水腫及肺出血的死亡病例，及四百零五位兒童重症住院，這個事件對整個社會造成重大衝擊。從1998年至今，腸病毒的流行看似減緩，但在今年腸病毒似乎有捲土重來的跡象，一個快速、準確、有效的檢測方法對於臨床診斷與重症病患的判定就扮演著非常重要的角色。新建立的CODEHOP RT-snPCR分子檢驗方法在美國已逐漸取代傳統的病毒培養，成為新的標準方法，因此我們藉此研究評估該方法是否真能應用於臨床檢驗。病毒培養與鑑定目前仍是病毒實驗室診斷方法中的「黃金標準」，以細胞培養配合間接螢光抗體試驗或中和試驗來鑑定病毒血清型，而聚合酶連鎖反應（PCR）漸漸成為一般實驗室快速鑑定腸病毒的檢驗方法。在本研究中，我們收集的2007年6-12月份204件臨床醫師懷疑腸病毒感染的臨床檢體，同時利用傳統的病毒培養與RT-snPCR加以測試，以評估RT-snPCR的可行性。在204件的檢體中，病毒培養呈現腸病毒陽性的件數為46，陽性率為23％，平均約需7到10個工作天；而RT-snPCR分子檢驗方法呈現陽性的件數為為115，陽性率為56％，CODEHOP鑑定出的種類，涵蓋範圍較病毒培養多，平均約需4到5個工作天，由此結果顯示CODEHOP方式較傳統培養的敏感性高出許多，也能在檢驗上縮短不少的時間。根據以上結果，在腸病毒流行的季節中，CODEHOP RT-snPCR檢驗方法的應用定能提供一個準確、快速且敏感的結果以供臨床醫師的處理。
並列摘要
Enterovirus infection is a common disease worldwide with an annual global incidence of over 5 billion, causing undifferentiated febrile illness, rash, abdominal symptoms, aseptic meningitis, paralysis, or fatal septicemia of neonates. The Enterovirus 71 outbreak in Taiwan during 1998 has caused 405 seriously ill cases which needed intensive care, together with 78 mortalities. Apart from a great impact on the affected families and the society, this tragedy has raised an enormous and intensive discussion on the epidemic prevention or control of enteroviruses. Among versatile discussed issues, a fast, timely, and accurate detection of enteroviruses has been believed to be vital and beneficial for the clinical diagnosis and management of sick children. During this year, ten years afterward, data collected from local clinics has indicated that a outbreak may occur. A fast and accurate molecular diagnosis method becomes a urgent demand, therefore, this study aimed to evaluate the feasibility of CODEHOP RT-snPCR, which has been utilized in USA, for the detection of enterovirus, as compared with the conventional virus culture. Among 204 samples suspected for enterovirus infection and collected between June to December, 2007, 46 samples (23%) were determined to be positive for enterovirus infection with conventional virus culture detection while 115 samples (56%) were determined to be positive with CODEHOP method. In the aspect of labor and time consumption, CODEHOP also possessed a great deal of advantage over the conventional method. Therefore, CODEHOP RT-snPCR is a feasible approach for clinical detection of enterovirus, which may provide more timely and accurate results and is of great value for clinical judgment and management of enterovirus infection.