台灣中部地區輪狀病毒種系發生之分析研究

由於輪狀病毒為造成全世界兒童急性腸胃炎之主要病原體，因此為了更充實本地輪狀病毒的資料庫及流行病毒株的演化性，故我們設計了該研究計劃。我們的實驗是收集中台灣部份不同年份和季節的醫院檢體進行輪狀病毒A群、B群及C群的初步篩選，所使用的主要實驗技術採用具專一性很高的反轉錄與聚合酶連鎖反應方法放大樣本並將所得之片段序列加以定序、分析與比較。本實驗總共收集了自1996年6月至9月與2001年10月至2002年3月共81個糞便檢體，結果總共篩選得到10株輪狀病毒A群的病毒株；然而，對於輪狀病毒B群與C群的篩選並無發現。在樣本與相關序列比對分析結果方面，所得到的10株輪狀病毒A群的產物進行基因序列定序，並採用電腦軟體比較分析其核苷酸基因序列與氨基酸序列，得到這10株病毒株序列皆屬於輪狀病毒VP7之部分基因序列，然後利用G-typing PCR方法來確認所分離出樣本之血清型，將所得到的序列分為G1，G3，G9等三種血清型，其中CS96-40為G1血清型與自烏拉圭所分離出來的MVD9816序列最為相似(兩者相似度為97.15%)；CS02-01則為G3血清型與由日本所分離出來的98-B31序列最為相似(兩者相似度為98.93%)；而CS01-05，CS01-06，CS01-07，CS01-09，CS01-13，CS02-02，CS02-03，CS02-04 共8株病毒株則為G9血清型與已證實為G9血清型的相關病毒株都很相似(相似度96.85~99.88%)，其中CS02-04與由日本所分離出來的SP2737序列，兩者間相差最少(其兩者之相似度為99.88%)。我們發現此次實驗中所分離出G3與G9之病毒株為台灣地區第一次所發表的亞型，因此對於未來台灣在輪狀病毒疫苗的研發上有相當重要的參考性。

Rotaviruses are a major cause of gastroenteritis in children worldwide. Phylogenetic analysis of the partial VP7 gene of the human rotavirus in central Taiwan. Total 81 stool samples from cases of acute gastroenteritis, occurring in the hospital of the central Taiwan at 1996, 2002, 2003, were selected by polyacrylamide gel electrophoresis and reverse transcription-polymerase chain reaction (RT-PCR) for partial VP7 gene of the human rotavirus group A, B, C. Then confirmed by comparative nucleotide and deduced amino acid sequence analysis for the relative sequence. A total of 10 stool sample were selected by polyacrylamide gel electrophoresis. The sequence analysis of the gene encoding the major neutralization glycoprotein (VP7) was performed on ten human isolates of serotype 1,3,9 of rotavirus and their genetic variations were examined. The partial VP7 gene of nucleotide sequence and the deduced amino acid sequence of CS96-40 of G1 are similar to MVD9816 (identity rate 97.15%, from Uruquay)；The sequence of CS02-01 of G3 are similar to 98-B31 (identity rate 98.93%, from Japan)；The sequence of CS01-05, CS01-06, CS01-07, CS01-09, CS01-13, CS02-02, CS02-03, CS02-04 and established G9 rotaviruses sequence are very similar (identity rate 96.85% ~ 99.88%), especially CS02-04 and SP2737 (from Japan) the identity rate is 99.88%. The first data of the human rotavirus group A serotype G3, G9 distribution in central Taiwan is reported. These data are important for the development of an appropriate rotavirus vaccine.