- 學位論文
台灣地區小兒急性腸胃炎症狀相關之腸道毒分子調查
MOLECULAR INVESTIGATION OF ACUTE GASTROENTERITIS SYNDROME RELATED ENTERIC VIRUSES AMONG CHILDREN IN TAIWAN
摘要
摘 要 輪狀病毒，腸腺病毒，杯狀病毒和腸病毒是引發急性腸胃炎相關的病毒。 這些腸胃炎病毒在台灣地區所扮演的角色仍是未知的。而且，在台灣7月到9月是盛行期也是輪狀病毒與腸病毒對於幼小孩童會造成混合感染的時期。 因此，本研究的目的是以分子生物學的方法(RT-PCR和ELISA)調查輪狀病毒與腸病毒在台灣的交叉流行期分布。 在台灣的腸病毒感染盛行期間，從全台各地的臨床實驗室總共收集了88個糞便檢體。每個檢體都先經過輪狀病毒的ELISA kit測試，然後再以RT-PCR來確認輪狀病毒與腸病毒的型別。 在88個具有腹瀉症狀的孩童糞便檢體中，有32個檢體呈現腸病毒陽性(佔36.4%) ，10個檢體呈現輪狀病毒陽性(佔11.4%)，而其中有3個檢體是具有兩種腸內病毒的混合感染(佔3.4%)。根據台灣的地理環境來觀察，輪狀病毒和腸病毒在北台灣較為流行，分別檢測出7個和21個陽性檢體；之後是中台灣，分別檢測出3個和10個陽性檢體；然後是南台灣，只有檢測出ㄧ個腸病毒的陽性檢體，輪狀病毒則無陽性反應。所有經輪狀病毒ELISA kit偵測所得到的陽性檢體之結果與輪狀病毒的RT-PCR結果相互吻合，證實RT-PCR之可行性。而且，在兩種方法之間沒有顯著的統計上的差異。 因此，證實RT-PCR在臨床糞便檢體之輪狀病毒檢測上，具有相同的效率。 在腸病毒的檢測中，以腸病毒71型的檢出率最高(佔47%)，其次為Coxsackievirus A(簡稱CA)的CA16 (佔38%)；而CA6與Echovirus(簡稱ECV) 的ECV 11分別檢測出兩個陽性檢體，而在本實驗中也檢測出1個檢體隸屬CA5型。在輪狀病毒的檢測中，將其陽性結果之基因序列與輪狀病毒的VP 7基因序列進行比對，其中有7個檢體的比對結果為輪狀病毒G3血清型，其相似度分別達98-99%；其餘檢體分別比對出2個G1與1個G2血清型別，其相似度分別達91-99%不等。以親緣演化的分析可發現其輪狀病毒的發生感染都在東亞地區(中國，日本和台灣)作循環。
並列摘要
ABSTART The etiologic agents related to acute gastroenteritis are rotavirus, enteric adenovirus, calicivirus, and enterovirus. It is still unknown what the role of these gastroenteritis viruses in Taiwan is. Besides, there were one peak season (July-September) regarded as mix infection for enterovirus and rotavirus for younger children population in Taiwan. Therefore, the object of this study is to investigate the prevalence of both enteric virus in Taiwan by molecular method (RT-PCR and ELISA). Total 88 fecal samples were collected from clinical laboratories distributed in different parts of Taiwan during enterovirus outbreak period in Taiwan. Each sample was first detected by rotavirus specific ELISA kit, and then the molecular typing of enterovirus and rotavirus was confirmed by reverse transcription-PCR. Among 88 fecal specimens collected from diarrheic children, 32 (36.4%) were positive for enterovirus, 10(11.4%) were for rotavirus and 3 (3.4%) patients were presented mix infection for both two enteric virus. According to geographic distribution, enterovirus and rotavirus were more prevalent in Northern Taiwan (21,7), followed by Central Taiwan (10,3) and then Southern Taiwan (1,0). All positive samples for rotavirus ELISA assay were also showed as positive for RT-PCR method. Besides, there was no significant statistical difference between two method. Therefore, RT-PCR method could perform same efficiency for detection of rotavirus in clinical stool samples. The most prevalent genotype were Enterovirus 71 (47 %), followed by coxsackievirus A 16(38%), and then CA6 and Ecovirus 11 were detected in two sample. We also find one sample was identified as CA5. Comparison of the nucleotide sequence with the VP7 gene of these rotavirus strains revealed that seven strains resembled closely with G3 serotype, the homology being 98 and 99 %, respectively. The following genotype was G1 for two samples and G2 for only one sample, shared 91-99 %. Phylogenetic analysis suggested that rotavirus were circulated in East Asia region (China, Japan and Taiwan).