Kariwa H, Kamimura M, Arikawa J, Yoshimatsu K, Takashima I, Hashimoto N
Department of Veterinary Public Health, Faculty of Veterinary Medicine, Hokkaido, Japan.
Microbiol Immunol. 1995;39(1):35-41. doi: 10.1111/j.1348-0421.1995.tb02165.x.
A polymerase chain reaction (PCR) for the detection of hantavirus genome was established and applied to analyze the mode of infection of Hantaan virus in adult ICR mice. The cDNA for the S genome segment of Hantaan virus was reverse-transcribed from the total RNA of organs of the infected mice. The sequence in the S genome segment of Hantaan virus was successfully amplified by reverse transcriptase (RT)-PCR followed by nested PCR. In 5-week-old ICR mice inoculated intraperitoneally with Hantaan virus, strain 76-118 (1.3 x 10(5) FFU/mouse), the virus was detected in clots and lungs from 3 to 10 days post-inoculation (p.i.) by nested PCR and virus-isolation techniques. No virus was detected in any specimens collected on 1 day and after 28 days p.i., and in spleens and brains through the observation period by both methods. The antibody which was measured by indirect immunofluorescence antibody assay (IFA) appeared at 7 days p.i. and the geometric mean titer was elevated to its maximum level of 1:203 at 10 days p.i., maintaining the same level until 35 days p.i. These results suggest that adult mice are transiently infected with Hantaan virus.
建立了一种用于检测汉坦病毒基因组的聚合酶链反应（PCR），并将其应用于分析成年ICR小鼠中汉滩病毒的感染模式。从感染小鼠器官的总RNA逆转录出汉滩病毒S基因组片段的cDNA。通过逆转录酶（RT）-PCR随后进行巢式PCR成功扩增了汉滩病毒S基因组片段中的序列。在腹腔接种汉滩病毒76-118株（1.3×10⁵ FFU/小鼠）的5周龄ICR小鼠中，通过巢式PCR和病毒分离技术在接种后3至10天（p.i.）在血凝块和肺中检测到病毒。在接种后1天和28天后收集的任何标本中，以及在整个观察期通过两种方法在脾脏和大脑中均未检测到病毒。通过间接免疫荧光抗体试验（IFA）检测的抗体在接种后7天出现，几何平均滴度在接种后10天升高到最高水平1:203，并在接种后35天保持相同水平。这些结果表明成年小鼠被汉滩病毒短暂感染。
