Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR.
Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR.
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DOI：
10.3389/fmicb.2013.00298
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发表时间：
2013
影响因子：
5.2
通讯作者：
Tsunetsugu-Yokota Y
中科院分区：
文献类型：
--
作者：
Ikeno S;Suzuki MO;Muhsen M;Ishige M;Kobayashi-Ishihara M;Ohno S;Takeda M;Nakayama T;Morikawa Y;Terahara K;Okada S;Takeyama H;Tsunetsugu-Yokota Y
麻疹减毒活病毒（MV）长期以来被认为是一种安全有效的疫苗，它已成为开发各种MV疫苗的基础。然而，由于MV是一种嗜人病毒，基于MV的疫苗的评价因缺乏小动物模型而受到阻碍。人源化小鼠是最近开发的一种系统，其中免疫缺陷小鼠移植有人胎儿组织或造血干细胞，可以代表合适的模型。在这里，我们开发了一种灵敏的一步定量逆转录（qRT）-PCR，同时测量核衣壳（N）和人RNase P mRNA水平。结果可用于监测人源化小鼠模型中的MV感染。使用这种方法，我们阐明了复制动力学的MV表达增强绿色荧光蛋白在体外和人源化小鼠中的流式细胞仪分析平行。由于我们的qRT-PCR系统足够灵敏，可以使用从少量细胞中提取的RNA检测MV表达，因此它可以用于通过连续采血监测人源化小鼠中的MV感染。
Live attenuated measles virus (MV) has long been recognized as a safe and effective vaccine, and it has served as the basis for development of various MV-based vaccines. However, because MV is a human-tropic virus, the evaluation of MV-based vaccines has been hampered by the lack of a small-animal model. The humanized mouse, a recently developed system in which an immunodeficient mouse is transplanted with human fetal tissues or hematopoietic stem cells, may represent a suitable model. Here, we developed a sensitive one-step quantitative reverse transcription (qRT)-PCR that simultaneously measures nucleocapsid (N) and human RNase P mRNA levels. The results can be used to monitor MV infection in a humanized mouse model. Using this method, we elucidated the replication kinetics of MV expressing enhanced green fluorescent protein both in vitro and in humanized mice in parallel with flow-cytometric analysis. Because our qRT-PCR system was sensitive enough to detect MV expression using RNA extracted from a small number of cells, it can be used to monitor MV infection in humanized mice by sequential blood sampling.
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影响因子：
5.2
作者：
Kato SI;Nagata K;Takeuchi K
通讯作者：
Takeuchi K
直肠内传播，全身感染和感染HIV-1的人型小鼠的CD4+ T细胞耗竭。
影响因子：
15.3
作者：
Sun, Zhifeng;Denton, Paul W.;Estes, Jacob D.;Othieno, Florence A.;Wei, Bangdong L.;Wege, Anja K.;Melkus, Michael W.;Padgett-Thomas, Angela;Zupancic, Mary;Haase, Ashley T.;Garcia, J. Victor
通讯作者：
Garcia, J. Victor
影响因子：
3.7
作者：
Akkina R
通讯作者：
Akkina R
影响因子：
5.4
作者：
Hashimoto, K;Ono, N;Yanagi, Y
通讯作者：
Yanagi, Y
影响因子：
3.1
作者：
Hummel, KB;Lowe, L;Rota, PA
通讯作者：
Rota, PA