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Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR
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 m...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2013
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3795360/ https://www.ncbi.nlm.nih.gov/pubmed/24130556 http://dx.doi.org/10.3389/fmicb.2013.00298 |
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author | Ikeno, Shota Suzuki, Moto-omi Muhsen, Mahmod Ishige, Masayuki Kobayashi-Ishihara, Mie Ohno, Shinji Takeda, Makoto Nakayama, Tetsuo Morikawa, Yuko Terahara, Kazutaka Okada, Seiji Takeyama, Haruko Tsunetsugu-Yokota, Yasuko |
author_facet | Ikeno, Shota Suzuki, Moto-omi Muhsen, Mahmod Ishige, Masayuki Kobayashi-Ishihara, Mie Ohno, Shinji Takeda, Makoto Nakayama, Tetsuo Morikawa, Yuko Terahara, Kazutaka Okada, Seiji Takeyama, Haruko Tsunetsugu-Yokota, Yasuko |
author_sort | Ikeno, Shota |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3795360 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-37953602013-10-15 Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR Ikeno, Shota Suzuki, Moto-omi Muhsen, Mahmod Ishige, Masayuki Kobayashi-Ishihara, Mie Ohno, Shinji Takeda, Makoto Nakayama, Tetsuo Morikawa, Yuko Terahara, Kazutaka Okada, Seiji Takeyama, Haruko Tsunetsugu-Yokota, Yasuko Front Microbiol Microbiology 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. Frontiers Media S.A. 2013-10-11 /pmc/articles/PMC3795360/ /pubmed/24130556 http://dx.doi.org/10.3389/fmicb.2013.00298 Text en Copyright © Ikeno, Suzuki, Muhsen, Ishige, Kobayashi-Ishihara, Ohno, Takeda, Nakayama, Morikawa, Terahara, Okada, Takeyama and Tsunetsugu-Yokota. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Ikeno, Shota Suzuki, Moto-omi Muhsen, Mahmod Ishige, Masayuki Kobayashi-Ishihara, Mie Ohno, Shinji Takeda, Makoto Nakayama, Tetsuo Morikawa, Yuko Terahara, Kazutaka Okada, Seiji Takeyama, Haruko Tsunetsugu-Yokota, Yasuko Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR |
title | Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR |
title_full | Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR |
title_fullStr | Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR |
title_full_unstemmed | Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR |
title_short | Sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative RT-PCR |
title_sort | sensitive detection of measles virus infection in the blood and tissues of humanized mouse by one-step quantitative rt-pcr |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3795360/ https://www.ncbi.nlm.nih.gov/pubmed/24130556 http://dx.doi.org/10.3389/fmicb.2013.00298 |
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