State Key Laboratory of Virology, Institute of Medical Virology, School of Medicine, Wuhan University, Wuhan, PR China.
Intervirology. 2013;56(3):172-7. doi: 10.1159/000345444. Epub 2013 Jan 9.
Hantaan virus (HTNV) is one of the main etiologic agents for hemorrhagic fever with renal syndrome in China. However, it is very difficult to isolate the virus from its original host, which hampers the viral characterization. This study describes an efficient method for isolating HTNV in suckling mice.
Hantavirus-infected Apodemus agrarius were screened by quantitative real-time PCR. The homogenates of one positive rodent lung tissue were inoculated into suckling mice for virus propagation through serial passages.
During the three passages in suckling mice, the number of viral RNA copies/nanogram of GAPDH mRNA increased significantly ranging from 477 to 7,278 and 46 to 4,898 in the tissues of brain and lung, respectively. Hantaviral antigens could be detected by indirect immunofluorescence assay and around 100-nm virion-like structures were also observed in brain tissue by transmission electron microscopy. No nucleotide exchange was found except for one in the 3'-non-coding domain of S segment when comparing the complete genome sequences from hantavirus in the first and the third passages.
These results suggest inoculation of suckling mice with suspected hantavirus-infected rodent samples is an efficient method for isolation and maintenance of HTNV.
汉坦病毒（HTNV）是中国引起肾综合征出血热的主要病原体之一。然而，从其自然宿主中分离病毒非常困难，这阻碍了病毒的特征描述。本研究描述了一种在乳鼠中分离 HTNV 的有效方法。
通过实时定量 PCR 筛选感染汉坦病毒的黑线姬鼠。将一个阳性啮齿动物肺组织的匀浆接种到乳鼠中，通过连续传代来进行病毒增殖。
在乳鼠中传代的 3 次过程中，脑和肺组织中病毒 RNA 拷贝数/ GAPDH mRNA 的纳米克数分别从 477 增加到 7278 和从 46 增加到 4898。通过间接免疫荧光测定可检测到汉坦病毒抗原，并且通过透射电子显微镜也可观察到脑组织中的约 100nm 大小的病毒样结构。与第一代和第三代传代的 S 片段 3'非编码区的一个核苷酸交换不同，除了 S 片段 3'非编码区的一个核苷酸交换外，比较汉坦病毒的全基因组序列时未发现其他核苷酸交换。
这些结果表明，接种乳鼠疑似感染汉坦病毒的啮齿动物样本是分离和维持 HTNV 的有效方法。
