荧光定量PCR检测感染登革2型病毒C6/36细胞和埃及伊蚊复制动态研究

摘要

To elucidate the dynamics of dengue virus in mosquito cells (C6/36) and Aedes egypti,FQ-PCR targeting on dengue virus type 2 was established to detect and monitor the viral load in C6/36 cells for 1-7 days post infection and that in Ae.egypti 1-13 days post-infection with an interval of 2 days.The proliferation of dengue virus in the mosquito cells changes regularly with time,more slowly in the first two days,then increased rapidly and reached the peak on the 3rd day but decreased slightly on the 6th day post-infection.While the proliferation of dengue virus in Ae.egypti began to increase on the 7th day and reached the peak on the 9th day,then fluctuated and decreased on the 13th day post infection.The results reflected that the reasonable time to harvest virus from C6/36 cells and from Ae.egypti might be 3rd day and 9th day post infection respectively.%本研究采用实时荧光定量PCR (FQ-PCR)方法,检测感染登革病毒C6/36细胞1～7d及感染埃及伊蚊1、3、5、7、9、11、13 d后,病毒在细胞和蚊虫体内增殖的动态变化.结果显示,随着时间增加,登革病毒增殖出现规律性变化.登革病毒在感染细胞1～2d后增殖不明显,3d后病毒开始大量复制达到高峰,但6d后有所下降.在蚊虫体内1～7d病毒增殖不明显,9d开始快速复制达到高峰,但11～13 d后下降至病毒载量稳定.实验表明登革病毒在细胞内复制增殖的最佳时间在接种病毒3d后,在蚊虫体内9d开始快速复制达到高峰增殖,到11 ～13 d后蚊虫体内病毒增殖稳定波动.