Frontiers in Microbiology (Oct 2013)

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

  • Shota eIkeno,
  • Shota eIkeno,
  • Moto-omi eSuzuki,
  • Moto-omi eSuzuki,
  • Mahmod eMuhsen,
  • Masayuki eIshige,
  • Mie eKobayashi,
  • Shinji eOhno,
  • Makoto eTakeda,
  • Tetsuo eNakayama,
  • Yuko eMorikawa,
  • Kazutaka eTerahara,
  • Seiji eOkada,
  • Haruko eTakeyama,
  • Haruko eTakeyama,
  • Yasuko Tsunetsugu Yokota

DOI
https://doi.org/10.3389/fmicb.2013.00298
Journal volume & issue
Vol. 4

Abstract

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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.

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