Scott T W, Hildreth S W, Beaty B J
Am J Trop Med Hyg. 1984 Mar;33(2):300-10. doi: 10.4269/ajtmh.1984.33.300.
The timing and sequence of eastern equine encephalitis (EEE) virus replication was studied in the organs from a colony strain of orally infected Culiseta melanura. Three methods of virus assay were used: fluorescent antibody (FA) staining of dissected organs; virus titration in cell culture of whole mosquitoes, dissected organs, hemolymph, and egg rafts; and transmission electron microscopy (TEM) of infected hindguts. EEE virus replicated rapidly in Cs. melanura, first in the posterior midgut, after which it disseminated into the hemocoel where hemolymph transported virus to other organs causing a systemic infection that eventually involved all organs examined, except ovarioles. No initial decrease in virus titer of whole mosquitoes or dissected organs was observed when mosquitoes were collected at daily intervals. Muscle tissue contained the greatest amount of specific fluorescence and the largest aggregates of virus that were visible by TEM. Dissemination of virus occurred rapidly, in some mosquitoes after less than or equal to 17 hours of extrinsic incubation (EI). All infected mosquitoes had disseminated infections after 3 days of EI. Maximum amounts of virus were obtained from whole mosquitoes on the 7th day of EI. FA staining of hindguts was determined to be a rapid and reliable method for detection of EEE virus dissemination and replication in Cs. melanura.
对来自口服感染黑尾库蚊群体品系的器官中东部马脑炎（EEE）病毒的复制时间和顺序进行了研究。使用了三种病毒检测方法：对解剖后的器官进行荧光抗体（FA）染色；对完整蚊子、解剖后的器官、血淋巴和卵筏进行细胞培养中的病毒滴定；以及对受感染的后肠进行透射电子显微镜（TEM）观察。EEE病毒在黑尾库蚊中迅速复制，首先在中肠后部，之后扩散到血腔，血淋巴将病毒运输到其他器官，引发全身感染，最终累及除卵巢小管外的所有检查器官。当每天间隔收集蚊子时，未观察到完整蚊子或解剖后器官的病毒滴度最初下降。肌肉组织含有最多的特异性荧光以及通过TEM可见的最大病毒聚集体。病毒传播迅速，在一些蚊子中，外在潜伏期（EI）小于或等于17小时后就发生了传播。所有受感染的蚊子在EI 3天后都出现了播散性感染。在EI第7天从完整蚊子中获得的病毒量最多。确定后肠的FA染色是检测EEE病毒在黑尾库蚊中传播和复制的一种快速且可靠的方法。
