臺灣博碩士論文加值系統

2009年四月，美國、墨西哥爆發了一波新的流行性感冒疫情，並且在全世界造成大流行；而此次的流行性感冒病毒經確認為一株新的A型H1N1流感病毒。A型流行性感冒病毒擁有八段基因，可分別轉錄出不同蛋白，其中血球凝集素（HA）與神經胺酸酶（NA）是最重要的膜蛋白，與病毒感染宿主細胞有關。流感病毒的基因會因抗原轉型（antigen shift）而造成重組，產生新型病毒，且大部分的人類對新型流感病毒都不具有免疫能力。鑒於以往SARS流行時醫護人員的感染機率較一般人為高，本研究利用血清學反應，比較台北榮民總醫院之醫護人員與一般民眾的血清抗體盛行率（seroprevalence rate），評估新型流感的流行對於醫謢人員之影響。利用血球凝集抑制試驗分析295個醫謢人員檢體與244個一般民眾檢體，醫護人員的血清陽性率明顯高於一般民眾，顯示其感染的危險性較高。在醫護人員中又依其接觸危險因子的機會，區分為高危險族群（Group 1）與低危險族群（Group 2）。高危險族群之血清盛行率也明顯高於低危險族群（30.8%；12.6%。p<0.05）。此外，本研究也收集四個時間點，分別為2009年9月、11月，以及2010年1月與4月之一般民眾檢體，探討新型流感在一般民眾之間的血清抗體盛行率在不同時間之變化。四個時間點之血清陽性率分別為：3.2%、2.9%、7.2%、0.8%；其中一月份的血清盛行率明顯高於其他月份（7.2%，p<0.05）。
　　台灣在2009年12月施行了全民施打H1N1新型流感疫苗的計畫，本研究也針對68名醫護人員收集其施打前後之血清，計算出其施打H1N1新型流感疫苗的血清抗體轉換率為10.3%。是否因樣本數過少或收案時間之影響，仍需進一步探討。

A new influenza outbreak started from Mexico and United States in April, 2009 and became pandemic. The origin of this pandemic influenza virus was found to be a novel type A H1N1 reassortment strain. The influenza A virus genomes comprise eight negative-strained RNA, and encode various proteins with different functions. For influenza A virus, attachment infectivity hemagglutinin (HA) and neuraminidase (NA) proteins are important surface glycoproteins encoded by segment 4 and 6. The error-prone RNA-dependent RNA polymerase of influenza virus and the gene rearrangement nature enables influenza virus to undergo antigenic drift and shift, resuiting in loss of immunity in the general population. Based on the experience of SARS epidemic, hospital staff has a higher risk of being infected. To understand the immune states of hospital staff in this new pandemic influenza, this study conducted a seroprevalence survey for the pandemic H1N1 influenza virus among hospital staff and general population. Adopting hemagglutinin-inhibition assay, we measured 295 serum samples from hospital staff with various contact risk and 244 control serum samples. The seroprevalece rate of hospital staff was significantly higher than control group. In addition, the seroprevalence of the hospital staff with higher contact risk was significantly higher than that of the hospital staff with lower contact risk (30.8%；12.6%。p&lt;0.05). The seropositive rates of the four time points (Sep. 2009；Nov. 2009；Jan. 2010；Apr. 2010) were 3.2%，2.9%，7.2% and 0.8%, respectively. Among them, the seropositive rate of Jan. 2010 time point was significantly higher than the others (p&lt;0.05).
　　The policy of pandemic H1N1 influenza vaccination was implemented in December, 2009, and this study collected 68 paired serum samples from pre-vaccination and post- vaccination hospital staff to analyze the seroconverstion rate. However, the seroconversion rate of pandemic H1N1 vaccine was 10.5%. Further studies may be required to determine whether the low seroconversion rate is influenced by small sample size.

中文摘要 i
Abstrate ii
縮寫表 iv
目錄 v
第一章　緒論 1
第一節 流行性感冒病毒 1
一、 歷史 1
二、 基本特性 2
三、 病毒感染與複製 4
四、 流行概況 6
五、 臨床病徵 7
六、 診斷 8
七、 預防與治療 10
第二節 實驗緣由與研究設計 13
一、 動機 13
二、 目的 13
三、　　研究設計 13
第二章　研究材料與方法 15
第一節　檢體來源 15
第二節　檢體前處理 15
第三節　病毒培養 17
第四節　血球凝集抑制試驗（HAI） 20
第五節　統計分析 20
第三章　實驗結果 22
第一節　比較醫護人員與一般民眾之血清盛行率 22
第二節　不同時間之血清盛行率變化 23
第三節　施打疫苗前後之血清轉換率 24
第四章　討論與結論 25
第一節　實驗結果討論 25
第二節　未來研究方向 28
參考文獻 30
圖表 36

表目錄
表一、H1N1新型流行性感冒病毒之RNA片段 36
表二、醫護人員與體檢民眾（control group）之間的血清盛行率 37
表三、不同時期之血清盛行率 38
表四、施打新型流感疫苗之血清轉換率 39
表五、real-time RT-PCR所用之Primer sets與probe 40
表六、real-time RT-PCR之反應條件。儀器：Smart-Cycler 41
表七、In-House RT-PCR之反應條件 41
表八、未注射H1N1新型流感疫苗問卷調查表 42
表九、已注射H1N1新型流感疫苗問卷調查表 43

圖目錄
圖一、MDCK細胞生長情形 44
圖二、以流感病毒感染MDCK後之細胞生長情形 45
圖三、病毒效價測試：血球凝集試驗（HA） 46
圖四、血清抗體效價測試：血球凝集抑制試驗（HAI） 47
圖五、以In-House RT-PCR偵測病毒之電泳結果 48
圖六、real-time RT-PCR偵測H1N1新型病毒之結果。儀器：Smart-Cycler 49
圖七、RT-PCR產物之定序結果 50
圖八、台灣新型流感之流行概況 52

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