A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies

Abstract Several xenogenic cell-based therapeutic products are currently under development around the world for the treatment of human diseases. Porcine islet cell products for treating human diabetes are a typical example. Since porcine cells possess endogenous retrovirus (PERV), which can replicat...

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Autores principales:	Ken Kono, Kiyoko Kataoka, Yuzhe Yuan, Keisuke Yusa, Kazuhisa Uchida, Yoji Sato
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b4ce8d8a941048a3b91f1d31adb3cd5c
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spelling	oai:doaj.org-article:b4ce8d8a941048a3b91f1d31adb3cd5c2021-12-02T12:03:15ZA highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies10.1038/s41598-020-78890-22045-2322https://doaj.org/article/b4ce8d8a941048a3b91f1d31adb3cd5c2020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78890-2https://doaj.org/toc/2045-2322Abstract Several xenogenic cell-based therapeutic products are currently under development around the world for the treatment of human diseases. Porcine islet cell products for treating human diabetes are a typical example. Since porcine cells possess endogenous retrovirus (PERV), which can replicate in human cells in vitro, the potential transmission of PERV has raised concerns in the development of these products. Four subgroups of infectious PERV have been identified, namely PERV-A, -B, -C, and recombinant PERV-A/C. Among them, PERV-A/C shows a high titre and there was a paper reported that an incidence of PERV-A/C viremia was increased in diseased pigs; thus, it would be important to monitor the emergence of PERV-A/C after transplantation of porcine products. In this study, we developed a highly sensitive method for the detection of PERV-A/C using next generation sequencing (NGS) technologies. A model PERV-C spiked with various doses of PERV-A/C were amplified by RT-PCR and the amplicons were analysed by NGS. We found that the NGS analysis allowed the detection of PERV-A/C at the abundance ratios of 1% and 0.1% with true positive rates of 100% and 57%, respectively, indicating that it would be useful for the rapid detection of PERV-A/C emergence after transplantation of porcine products.Ken KonoKiyoko KataokaYuzhe YuanKeisuke YusaKazuhisa UchidaYoji SatoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-8 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ken Kono Kiyoko Kataoka Yuzhe Yuan Keisuke Yusa Kazuhisa Uchida Yoji Sato A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies
description	Abstract Several xenogenic cell-based therapeutic products are currently under development around the world for the treatment of human diseases. Porcine islet cell products for treating human diabetes are a typical example. Since porcine cells possess endogenous retrovirus (PERV), which can replicate in human cells in vitro, the potential transmission of PERV has raised concerns in the development of these products. Four subgroups of infectious PERV have been identified, namely PERV-A, -B, -C, and recombinant PERV-A/C. Among them, PERV-A/C shows a high titre and there was a paper reported that an incidence of PERV-A/C viremia was increased in diseased pigs; thus, it would be important to monitor the emergence of PERV-A/C after transplantation of porcine products. In this study, we developed a highly sensitive method for the detection of PERV-A/C using next generation sequencing (NGS) technologies. A model PERV-C spiked with various doses of PERV-A/C were amplified by RT-PCR and the amplicons were analysed by NGS. We found that the NGS analysis allowed the detection of PERV-A/C at the abundance ratios of 1% and 0.1% with true positive rates of 100% and 57%, respectively, indicating that it would be useful for the rapid detection of PERV-A/C emergence after transplantation of porcine products.
format	article
author	Ken Kono Kiyoko Kataoka Yuzhe Yuan Keisuke Yusa Kazuhisa Uchida Yoji Sato
author_facet	Ken Kono Kiyoko Kataoka Yuzhe Yuan Keisuke Yusa Kazuhisa Uchida Yoji Sato
author_sort	Ken Kono
title	A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies
title_short	A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies
title_full	A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies
title_fullStr	A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies
title_full_unstemmed	A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies
title_sort	highly sensitive method for the detection of recombinant perv-a/c env rna using next generation sequencing technologies
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/b4ce8d8a941048a3b91f1d31adb3cd5c
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A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies

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