title: Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR.
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.
authors.affiliation: Department of Immunology, National Institute of Infectious Diseases Tokyo, Japan ; Cooperative Major in Advanced Health Science, Tokyo University of Agriculture and Technology/Waseda University Graduate School of Collaborative Education Curriculum Tokyo, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Ikeno
authors.forename: Moto-Omi
authors.initials: MO
authors.lastname: Suzuki
authors.forename: Mahmod
authors.initials: M
authors.lastname: Muhsen
authors.forename: Masayuki
authors.initials: M
authors.lastname: Ishige
authors.forename: Mie
authors.initials: M
authors.lastname: Kobayashi-Ishihara
authors.forename: Shinji
authors.initials: S
authors.lastname: Ohno
authors.forename: Makoto
authors.initials: M
authors.lastname: Takeda
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nakayama
authors.forename: Yuko
authors.initials: Y
authors.lastname: Morikawa
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Terahara
authors.forename: Seiji
authors.initials: S
authors.lastname: Okada
authors.forename: Haruko
authors.initials: H
authors.lastname: Takeyama
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Tsunetsugu-Yokota
keywords: EGFP expression; flow cytometry; humanized mouse; measles virus infection; quantitative RT-PCR