Sensitive Detection of Measles Virus Infection in the Blood and Tissues of Humanized Mouse by One-step Quantitative RT-PCR

• Main Authors: Shota eIkeno, Moto-omi eSuzuki, Mahmod eMuhsen, Masayuki eIshige, Mie eKobayashi, Shinji eOhno, Makoto eTakeda, Tetsuo eNakayama, Yuko eMorikawa, Kazutaka eTerahara, Seiji eOkada, Haruko eTakeyama, Yasuko Tsunetsugu Yokota
• Format: Article
• Language: English
• Published: Frontiers Media S.A. 2013-10-01
• Series: Frontiers in Microbiology
• Subjects: Flow Cytometry; quantitative RT-PCR; measles virus infection; humanized mouse; EGFP expression
• Online Access: http://journal.frontiersin.org/Journal/10.3389/fmicb.2013.00298/full

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 EGFP 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.