高雄縣梓官鄉位於南台灣高B型、C型及D型感染的鄉鎮，於民國八十六年針對2909位鄉民年齡超過45歲之篩檢發現B型肝炎表面抗原 (HBsAg) 及C型肝炎抗體 (anti HCV) 盛行率分別為12.8% 及41.6%，而B型肝炎帶原者中其Anti-HDV呈陽性反應者佔15.3%。梓官鄉可分成沿海八村及或內陸七村，其沿海區域之盛行率為內陸之區域的兩倍 (61.4% v.s. 29.1%)，而基因型1b及基因型2a為兩大主要亞型。我們嘗試以親緣關係分析 (phylogenetic analysis) 去尋找造成這兩個相近區域有如此差異的原因並且假設其傳播路徑為沿海向內陸。除了參個村落僅取1b基因型 (genotype) 外，自這15個村各取一個基因型1b及一個基因型2a檢體，總共27人 (內羣：ingroup)，包括8位男性及19位女性，平均年齡54.7歲 (範圍: 45-70)，這27人均來自不同家庭。這些檢體以反轉錄 (Reverse transcriptase, RT) 及聚合鏈反應 (Polymerase chain reaction, PCR) 方式，接著再以重疊聚合鏈反應 (nested PCR) 方式來檢測C型肝炎病毒之NS5B 段落。自梓官鄉之外的南臺灣取10人當局部對照組 (local control)，另外自基因庫〔http://hcv.lanl.gov/content/hcv-db/index〕採取30個NS5B序列。以親緣關係樹 (phylogenetic tree) 分析可得同一鄉之鄉民各依其基因型呈兩大主要分群，呈現高度群集性的現象，但是並未發現如預期其傳播路徑為沿海向內陸抑或呈現同一基因型依其居住地而分成沿海或內陸兩大羣。而其內羣中來自不同村之C型肝炎病毒遺傳距離相近 (基因型1b及2a間最大及最小遺傳距離分別為0.0869 vs. 0.0098及0.0996 vs. 0.0334)，均可能是因為C型肝炎病毒於盛行區內散播之廣泛。此外，依據臺灣原住民過去與外來人種接觸史等，我們嘗試自基因庫上擷取不同國家之所有NS5B序列與梓官鄉民作遺傳距離分析發現梓官鄉之C型肝炎病毒基因型1b可能來自三個不同來源，且其顯示各國間C型肝炎病毒在此盛行區交流及廣泛之散播。

Tzukuan is an HBV-, HCV-, HDV- endemic township in southern Taiwan. Based on a mass screening on 2909 residents age of 45 years or more in 1997, the prevalence rates of HBsAg and anti-HCV were 12.8% and 41.6% respectively. Of HBsAg carriers, 15.3% were positive for anti-HDV. Tzukaun was divided into coastal area and inland area. The prevalence of anti-HCV of coastal area was two times higher than that of inland area (61.4% v.s. 29.1%) and genotype 1b and 2a are the main two subtypes. We wish to find the causes of discrepancy in these nearby areas by phylogenetic analysis. Stratified by the living areas, coastal or inland, 27 samples were picked up (ingroup). HCV sequence of NS5B region could be detected by RT-PCR then a nested PCR in eight males and ninteen females with mean age of 54.8 years old (range: 45-70). None of these 27 residents came from the same family. Another 10 HCV infected persons whose living townships also in southern Taiwan but other than Tzukuan were enrolled as local controls. From GenBank, 30 different HCV isolates were included. Phylogenic analysis unequivocally confirmed the simultaneous spread of two different HCV strains in this township clusters according to their subtypes were noted. A trend of the spreading from coastal to land area or an ultra-aggregation phynomenon which according to their living area, as we suspected, were not noted between Tzukuan’s residents. In ingroup, the short genetic distance between the isolates of C hepatitis virus which came from different villages might be caused from the wide-spreading of HCV in this endemic area (the maximal and minimal genetic distance in 1b or 2a isolates are 0.0869 vs. 0.0098 and 0.0996 vs. 0.0334). Besides, according to the contacting history to foreigner by our aborigine tribes, from genebank, all isolates from different countries were included and three possible origins of HCV genotype 1b were noted in Tzukuan. All these findings might be caused from frequently HCV inflow in this endemic area and wide-spreading of HCV between different countries.

目錄．．．．．．．．．．．．．．．．．．．．．．．．．．．i
中文摘要．．．．．．．．．．．．．．．．．．．．．．．．．iv
英文摘要．．．．．．．．．．．．．．．．．．．．．．．．．vi
前言．．．．．．．．．．．．．．．．．．．．．．．．．．．1
研究背景．．．．．．．．．．．．．．．．．．．．．．．．．6
研究目的．．．．．．．．．．．．．．．．．．．．．．．．．7
材料及方法．．．．．．．．．．．．．．．．．．．．．．．．8
結果．．．．．．．．．．．．．．．．．．．．．．．．．．．18
討論．．．．．．．．．．．．．．．．．．．．．．．．．．23
結論．．．．．．．．．．．．．．．．．．．．．．．．．．．27
參考文獻．．．．．．．．．．．．．．．．．．．．．．．．．28
圖表．．．．．．．．．．．．．．．．．．．．．．．．．．35
表一、包含基因型1至6之第一羣基因庫NS5B序列．．．．．．35
表二、內羣、局部對照組及第一羣基因庫NS5B序列間之最大及最小遺傳距離．．．．．．．．．．．．．．．．．．．．．．．．．36
表三、梓官鄉C型肝炎病毒基因型1b之可能三大來源．．．．．．37
表四、基因型2a之內羣與第三羣基因庫NS5B序列間遺傳距離最近之前三者．．．．．．．．．．．．．．．．．．．．．．．．．．．38
圖一、以最簡約法 (Maximum Parsimony method) 分析之親緣關係樹．．．．．．．．．．．．．．．．．．．．．．．．．．．．39
圖二、以鄰接法 (Neighbor-joining method) 分析之親緣關係樹40
圖三、局部放大以鄰接法分析之親緣關係樹基因型1b之分枝．．．41
圖四、局部放大以鄰接法分析之親緣關係樹基因型2a之分枝及分型錯誤者．．．．．．．．．．．．．．．．．．．．．．．．．．．42
圖五、以鄰接法分析之親緣關係樹局部放大基因型1b之分枝並顯示距離．．．．．．．．．．．．．．．．．．．．．．．．．．．．43
圖六、以鄰接法分析之親緣關係樹局部放大基因型2a之分枝並顯示距離．．．．．．．．．．．．．．．．．．．．．．．．．．．．44
附錄．．．．．．．．．．．．．．．．．．．．．．．．．．．44
附錄一、C型肝炎病毒之基因結構．．．．．．．．．．．．．．45
附錄二、Inno Lipa kit 利用不同基因型之去氧核糖核酸探子與5’端非轉錄區上七個不同區段結合與否以判斷基因型之方式．．．．46
附錄三、基因庫基因 (M62321.1)、內羣及局部對照組之核酸序列．．．．．．．．．．．．．．．．．．．．．．．．．．．．47
附錄四、基因庫基因 (M62321.1)、內羣及局部對照組之蛋白質序列．．．．．．．．．．．．．．．．．．．．．．．．．．．．54
附錄五、內羣、局部對照組及第一羣基因庫NS5B序列間之遺傳距離 ．．．．．．．．．．．．．．．．．．．．．．．．．．．57
附錄六、HCV 1b之第二羣基因庫NS5B序列之目錄號碼．．．．．64
附錄七、HCV 2a之第三羣基因庫NS5B序列之目錄號碼．．．．．74

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李筱峰 台灣史100件大事 台北：玉山社出版社，民國八十八年。