AUTHOR=Ikeno Shota , Suzuki Moto-omi , Muhsen Mahmod , Ishige Masayuki , Kobayashi Mie , Ohno Shinji , Takeda Makoto , Nakayama Tetsuo , Morikawa Yuko , Terahara Kazutaka , Okada Seiji , Takeyama Haruko , Yokota Yasuko T. TITLE=Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR JOURNAL=Frontiers in Microbiology VOLUME=4 YEAR=2013 URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2013.00298 DOI=10.3389/fmicb.2013.00298 ISSN=1664-302X ABSTRACT=

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.