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Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients

Genetic testing using reverse transcriptase real-time polymerase chain reaction (rRT-PCR) is the mainstay of diagnosis of COVID-19. However, it has not been fully investigated whether infectious viruses are contained in SARS-CoV-2 genome-positive specimens examined using the rRT-PCR test. In this st...

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Autores principales: Igarashi, Emiko, Tani, Hideki, Tamura, Kosuke, Itamochi, Masae, Shimada, Takahisa, Saga, Yumiko, Inasaki, Noriko, Hasegawa, Sumiyo, Yazawa, Shunsuke, Sasajima, Hitoshi, Kaya, Hiroyasu, Nomura, Satoshi, Itoh, Hiroyuki, Takeda, Shin-ichi, Yamamoto, Yoshihiro, Yamashiro, Seiji, Ichikawa, Tomomi, Horie, Yukio, Hirota, Kojiro, Hirano, Norikazu, Kawashiri, Chikako, Oishi, Kazunori
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8578003/
https://www.ncbi.nlm.nih.gov/pubmed/34774431
http://dx.doi.org/10.1016/j.jiac.2021.10.028
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author Igarashi, Emiko
Tani, Hideki
Tamura, Kosuke
Itamochi, Masae
Shimada, Takahisa
Saga, Yumiko
Inasaki, Noriko
Hasegawa, Sumiyo
Yazawa, Shunsuke
Sasajima, Hitoshi
Kaya, Hiroyasu
Nomura, Satoshi
Itoh, Hiroyuki
Takeda, Shin-ichi
Yamamoto, Yoshihiro
Yamashiro, Seiji
Ichikawa, Tomomi
Horie, Yukio
Hirota, Kojiro
Hirano, Norikazu
Kawashiri, Chikako
Oishi, Kazunori
author_facet Igarashi, Emiko
Tani, Hideki
Tamura, Kosuke
Itamochi, Masae
Shimada, Takahisa
Saga, Yumiko
Inasaki, Noriko
Hasegawa, Sumiyo
Yazawa, Shunsuke
Sasajima, Hitoshi
Kaya, Hiroyasu
Nomura, Satoshi
Itoh, Hiroyuki
Takeda, Shin-ichi
Yamamoto, Yoshihiro
Yamashiro, Seiji
Ichikawa, Tomomi
Horie, Yukio
Hirota, Kojiro
Hirano, Norikazu
Kawashiri, Chikako
Oishi, Kazunori
author_sort Igarashi, Emiko
collection PubMed
description Genetic testing using reverse transcriptase real-time polymerase chain reaction (rRT-PCR) is the mainstay of diagnosis of COVID-19. However, it has not been fully investigated whether infectious viruses are contained in SARS-CoV-2 genome-positive specimens examined using the rRT-PCR test. In this study, we examined the correlation between the threshold Cycle (Ct) value obtained from the rRT-PCR test and virus isolation in cultured cells, using 533 consecutive clinical specimens of COVID-19 patients. The virus was isolated from specimens with a Ct value of less than 30 cycles, and the lower the Ct value, the more efficient the isolation rate. A cytopathic effect due to herpes simplex virus type 1 contamination was observed in one sample with a Ct value of 35 cycles. In a comparison of VeroE6/TMPRSS2 cells and VeroE6 cells used for virus isolation, VeroE6/TMPRSS2 cells isolated the virus 1.7 times more efficiently than VeroE6 cells. There was no significant difference between the two cells in the mean Ct value of the detectable sample. In conclusion, Lower Ct values in the PCR test were associated with higher virus isolation rates, and VeroE6/TMPRSS2 cells were able to isolate viruses more efficiently than VeroE6 cells.
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spelling pubmed-85780032021-11-10 Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients Igarashi, Emiko Tani, Hideki Tamura, Kosuke Itamochi, Masae Shimada, Takahisa Saga, Yumiko Inasaki, Noriko Hasegawa, Sumiyo Yazawa, Shunsuke Sasajima, Hitoshi Kaya, Hiroyasu Nomura, Satoshi Itoh, Hiroyuki Takeda, Shin-ichi Yamamoto, Yoshihiro Yamashiro, Seiji Ichikawa, Tomomi Horie, Yukio Hirota, Kojiro Hirano, Norikazu Kawashiri, Chikako Oishi, Kazunori J Infect Chemother Note Genetic testing using reverse transcriptase real-time polymerase chain reaction (rRT-PCR) is the mainstay of diagnosis of COVID-19. However, it has not been fully investigated whether infectious viruses are contained in SARS-CoV-2 genome-positive specimens examined using the rRT-PCR test. In this study, we examined the correlation between the threshold Cycle (Ct) value obtained from the rRT-PCR test and virus isolation in cultured cells, using 533 consecutive clinical specimens of COVID-19 patients. The virus was isolated from specimens with a Ct value of less than 30 cycles, and the lower the Ct value, the more efficient the isolation rate. A cytopathic effect due to herpes simplex virus type 1 contamination was observed in one sample with a Ct value of 35 cycles. In a comparison of VeroE6/TMPRSS2 cells and VeroE6 cells used for virus isolation, VeroE6/TMPRSS2 cells isolated the virus 1.7 times more efficiently than VeroE6 cells. There was no significant difference between the two cells in the mean Ct value of the detectable sample. In conclusion, Lower Ct values in the PCR test were associated with higher virus isolation rates, and VeroE6/TMPRSS2 cells were able to isolate viruses more efficiently than VeroE6 cells. Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. 2022-02 2021-11-10 /pmc/articles/PMC8578003/ /pubmed/34774431 http://dx.doi.org/10.1016/j.jiac.2021.10.028 Text en © 2021 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Note
Igarashi, Emiko
Tani, Hideki
Tamura, Kosuke
Itamochi, Masae
Shimada, Takahisa
Saga, Yumiko
Inasaki, Noriko
Hasegawa, Sumiyo
Yazawa, Shunsuke
Sasajima, Hitoshi
Kaya, Hiroyasu
Nomura, Satoshi
Itoh, Hiroyuki
Takeda, Shin-ichi
Yamamoto, Yoshihiro
Yamashiro, Seiji
Ichikawa, Tomomi
Horie, Yukio
Hirota, Kojiro
Hirano, Norikazu
Kawashiri, Chikako
Oishi, Kazunori
Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients
title Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients
title_full Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients
title_fullStr Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients
title_full_unstemmed Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients
title_short Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients
title_sort viral isolation analysis of sars-cov-2 from clinical specimens of covid-19 patients
topic Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8578003/
https://www.ncbi.nlm.nih.gov/pubmed/34774431
http://dx.doi.org/10.1016/j.jiac.2021.10.028
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