原位分子杂交检测大鼠肾综合征出血热病毒隐性感染和病毒RNA的体内分布

目的和方法：用异羟基洋地黄毒苷配基－１１－ｄＵＴＰ随机引物延伸法分别标记肾综合征出血热（ＨＦＲＳ）汉坦株病毒Ｍ与ＳＲＮＡ全片段ｃＤＮＡ作为探针，用核酸原位分子杂交法检测某动物所一批大鼠ＨＦＲＳ病毒的隐性感染及病毒ＲＮＡ的组织细胞定位和分布．结果：９８只大鼠中８８只病毒ＲＮＡ阳性，阳性率８９．７９％，阳性率最高的脏器是肝脏７９／８５，其次为心脏２２／８３，再次为肾脏１４／７９，其余脏器阳性率较低．阳性部位主要在各脏器的实质细胞胞浆内，呈细颗粒状，未见于核内．结论：结果说明本批大鼠ＨＦＲＳ病毒隐性感染率极高；大鼠ＨＦＲＳ病毒ＲＮＡ的阳性检出率明显高于同批大鼠病毒抗原和血清抗体的阳性检出率，说明原位分子杂交检测病毒ＲＮＡ对确定动物病毒隐性感染具有重要的流行病学意义．病毒ＲＮＡ主要存在于肝脏，提示肝脏可能是病毒的主要潜伏部位，病毒可能主要由粪便排除．

In situ hybridization detection of hemorrhagic fever with renal syn-drome virus persistent infection and viral RNA distribution in labora-tory rats

Objective and Methods : Digoxigenin-labelled Hantaan specific cDNA probes were used in thisstudy for in situ hybridization detection of hemorrhagic fever with renal syndrome(HFRS ) virus persistentinfection and viral RNA distribution in naturally infected laboratory rats. Results : Of 98 rats, 88 displayedpositive hybridization signals in one or more of their tissues. Viral RNA was mainly localized in the cytoplasmwith granular staining and distributed in hepatocytes, to some extent in myocardial cells, reticuloendothelialcells of lymphatic tissues, respiratory epithelial cells of lung, renal tubular cells, neurons and blood serum.Viral RNA was shown to have higher positive rate in livers than in other tissues and the rate was also higherthan that of viral antigens and serum antibody in the same group of rats in our early report. Conclusions :This results indicated that HFRS virus persistent infection rate in laboratory rats was much higher than previ-ously known, and only in part of viral RNA positive rats , could viral antigens and antibody be detected,therefore viral RNA detection for confirmation of persistent infection could be of important significance forepidemiological study of HFRS. The fact that viral RNA mainly existed in liver with higher positive ratesmight suggest that liver was the main organ habouring HFRS virus in persistent infected rats, and that thevirus might be excreted from liver via intestine into the environments.