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从实验性和自然感染小鼠中检测小鼠诺如病毒的分子方法。

Molecular detection of murine norovirus from experimentally and spontaneously infected mice.

作者信息

Goto Kazuo, Hayashimoto Nobuhito, Yasuda Masahiko, Ishida Tomoko, Kameda Shuko, Takakura Akira, Itoh Toshio

机构信息

Central Institute for Experimental Animals, Miyamae-ku, Kawasaki, kanagawa, Japan.

出版信息

Exp Anim. 2009 Apr;58(2):135-40. doi: 10.1538/expanim.58.135.

Abstract

The RNA polymerase gene of murine norovirus (MNV) was isolated from feces and organ samples by the reverse transcription (RT)-polymerase chain reaction (PCR). For experimental infection, homogenate of cecum obtained from an MNV-infected mouse was gavaged to 3 C.B-17-Prkdc(scid) (scid) mice and 3 ICR mice at 6 weeks of age. Sixty days after oral inoculation, MNV was isolated from the cecum (3/3 scid and 3/3 ICR), feces (3/3 scid and 3/3 ICR), duodenum (1/3 scid and 3/3 ICR), liver (1/3 scid and 1/3 ICR), and spleen (3/3 ICR) samples, but MNV was not detected in the brain, heart, kidney, lung, salivary gland, ovary, thymus, or uterus samples of any of the orally inoculated mice. Feces of males cohabiting with MNV infected dams were positive for viral RNA after 18 days of cohabitation, but 8 fetuses (embryonic day 18.0) derived from the dams were negative for the virus. The results suggest that the cecum and feces are the most suitable sample types for the detection of MNV in infected animals and that caesarean section is efficient for the elimination of the virus. In terms of spontaneous infection, the RNA polymerase gene of MNV was isolated from 33/245 (13.1%) cecum samples derived from 15/59 (25.4%) facilities, and the sequence analysis revealed that at least 5 types of the virus were prevalent. This is the first report on MNV infection in mouse colonies in Japan.

摘要

通过逆转录(RT)-聚合酶链反应(PCR)从小鼠粪便和器官样本中分离出鼠诺如病毒(MNV)的RNA聚合酶基因。为进行实验性感染,将从感染MNV的小鼠获得的盲肠匀浆灌胃给6周龄的3只C.B-17-Prkdc(scid)(scid)小鼠和3只ICR小鼠。口服接种60天后,从盲肠(3/3只scid小鼠和3/3只ICR小鼠)、粪便(3/3只scid小鼠和3/3只ICR小鼠)、十二指肠(1/3只scid小鼠和3/3只ICR小鼠)、肝脏(1/3只scid小鼠和1/3只ICR小鼠)和脾脏(3/3只ICR小鼠)样本中分离出MNV,但在任何口服接种小鼠的脑、心、肾、肺、唾液腺、卵巢、胸腺或子宫样本中均未检测到MNV。与感染MNV的母鼠同居的雄性小鼠粪便在同居18天后病毒RNA呈阳性,但来自这些母鼠的8只胎儿(胚胎第18.0天)病毒检测呈阴性。结果表明,盲肠和粪便是检测感染动物中MNV最合适的样本类型,剖腹产对于消除病毒是有效的。就自然感染而言,从来自15/59(25.4%)设施的245份盲肠样本中的33份(13.1%)分离出MNV的RNA聚合酶基因,序列分析显示至少有5种病毒流行。这是日本关于小鼠群体中MNV感染的首次报告。

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