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標題: Characterization the Infection of Japanese Encephalitis Virus Genotype Ⅰ and Ⅲ
作者: 廖淑吟
Shu-Ying Liao
關鍵字: 日本腦炎;基因型;感染;Japanese Encephalitis Virus;JEV;Genotype;Flavivirus
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Japanese encephalitis virus (JEV) is a member of the Flavivirus genus of the family of Flaviviridae. Since 1990s, the JEV genotype III replaced by genotype I had been indicated in several southeast Asia Countries. According to the virological surveillance and molecular epidemiology of JEV among pig farms in Taiwan, the results also shown that the dominant JEV genotype in Taiwan has switched from genotype Ⅲ to Ⅰduring 2008 to 2009. In Japan, after JEV genotype shift, the JE confirmed cases were decreased from 50~100 to less than 10 cases annually, and suggested the genotype I JEVs were less virulence, and then the JEV vaccination program was ceased in 2005. However, the decreasing of confirmed case was not observed after JEV genotype replacement in Taiwan and Thailand. Also, some studies also indicated that there were no significant differences between genotype I and III in mouse neurovirulence. Thus, the possible mechanism(s) of JEV genotype III replaced by genotype I and the pathogenicity of genotype I and III in human are unclear.
In this study, we estimate and compare the pathogenicity of genotype Ⅰ and Ⅲ by calculating the asymptomatic ratio and study infectivity of genotype I and III JEVs in different cell lines under different conditions. We collect the serum specimens during genotype Ⅰ(2010-2011) and genotype Ⅲ (1994 and 1999) endemic periods in Taiwan. The status of JEV infection was serologically determined using plaque reduction neutralization assay (PRNT) and IgM antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA). The asymptomatic ratio was estimated using JE confirmed cases as denominator and JEV infected cases as the numerator. The values were adjusted by JE uninfected cases with manifestation of encephalitis, gander and vaccination coverage using the log-linear binomial regression model. The results shown that the asymptomatic ratio of JEV GⅢ and GⅠ virus infection were 1:7,510±1,270 and 1:6,211±722.6, respectively. Based on the similar asymptomatic ratios of JEV GⅢ and GⅠ, we suggested the pathogenicity in human was similar between JEV genotype Ⅰ and Ⅲ virus. The replication of genotype I JEVs were better than genotype III viruses in DF-1, CER, DE and PK-15 cell lines, but not in C6/36 cells. Based on these results, we suggested that the genotype JEVs might have replication advantage in amplification hosts, but in vivo studies were needed.

日本腦炎病毒(Japanese encephalitis virus)屬於黃病毒科(Flaviviridae)、黃病毒屬(Flavivirus)之病毒。自1990 年代以來,許多東南亞國家流行的日本腦炎病毒發生由原本的第三型基因型轉變成以第一型基因型;台灣於2009~2010年間,利用對豬場與病媒蚊監測進行分子流行病學分析後,也發現於台灣亦已發生並完成基因型轉型之現象。日本在發生日本腦炎病毒基因型別轉變後,確定病例數由每年50~100例銳減至10例以下,並由此觀察結果此推論第一型基因型病毒致病能力較第三型基因型為弱,並且於2005年終止日本腦炎國家疫苗接種計畫。然而反觀台灣與泰國等其他國家,並未在發生日本腦炎病毒基因型轉型後觀察到日本腦炎確定病例下降之現象;且在小鼠之毒力試驗的相關研究中,亦無發現日本腦炎病毒不同基因型之毒力有顯著差異。僅就上述觀察及文獻之結果,日本腦炎病毒第一型取代第三型病毒機制,及此二基因型病毒在人類之致病力是否有所不同仍待研究。
本研究利用日本腦炎病毒第一型與第三型之不顯感染比例估算,以評估日本腦炎病毒第一型與第三型在人類之致病力是否有所不同;並利用不同條件感染於不同宿主細胞株之病毒生長曲線之進行比較,以探討在傳播途徑之不同宿主中日本腦炎第一基因型病毒取代第三型基因型病毒之可能機轉及。本研究中利用在第三基因型流行時所收集之血清檢體(1994、1999年)與第一基因型流行時所收集之血清檢體(2010-2011年)利用MAC-ELISA及PRNT50來估算該地區日本腦炎盛行率並計算其不顯感染比例(確定病例數比被感染數),並針對背景症狀、性別、疫苗接種情況利用Log-linear binomial regression model進行校正,得到台灣日本腦炎病毒之不顯感染比例第三型與第一型分別為1:7,510±1,270 與 1:6,211±722.6,發現以此不顯感染比例比較第一基因型與第三型基因型病毒在感染台灣居民後之致病力並無顯著差異。而病毒生長曲線部分,在禽類細胞DF-1、CER、DE細胞株中皆可以觀察到第一基因型日本腦炎病毒複製情形優於部分第三型基因型病毒,在以高溫培養的豬隻PK-15細胞株中亦發現一基因型日本腦炎病毒複製情形優於部分第三型基因型病毒之情形。但是在白線斑蚊C6/36細胞株則發現不論在不同溫度或是感染劑量之條件皆無顯著差異。由此結果推論,在日本腦炎第一基因型病毒取代第三型基因型病毒之可能機轉中,日本腦炎第一型與第三型基因型病毒於增幅宿主(豬及鳥類)中之適應性與複製能力之差異可能為重要之原因之一,但仍須進一步進行動物實驗。
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