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Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety

BACKGROUND & OBJECTIVES: Polio, measles, rubella, influenza and rotavirus surveillance programmes are of great public health importance globally. Virus isolation using cell culture is an integral part of such programmes. Possibility of unintended isolation of SARS-CoV-2 from clinical specimens p...

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Autores principales: Pawar, Shailesh D., Kode, Sadhana S., Keng, Sachin S., Tare, Deeksha S., Diop, Ousmane M., Abraham, Priya, Sharma, Deepa K., Sangal, Lucky, Yadav, Pragya D., Potdar, Varsha A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Wolters Kluwer - Medknow 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9552391/
https://www.ncbi.nlm.nih.gov/pubmed/35859439
http://dx.doi.org/10.4103/ijmr.ijmr_1448_21
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author Pawar, Shailesh D.
Kode, Sadhana S.
Keng, Sachin S.
Tare, Deeksha S.
Diop, Ousmane M.
Abraham, Priya
Sharma, Deepa K.
Sangal, Lucky
Yadav, Pragya D.
Potdar, Varsha A.
author_facet Pawar, Shailesh D.
Kode, Sadhana S.
Keng, Sachin S.
Tare, Deeksha S.
Diop, Ousmane M.
Abraham, Priya
Sharma, Deepa K.
Sangal, Lucky
Yadav, Pragya D.
Potdar, Varsha A.
author_sort Pawar, Shailesh D.
collection PubMed
description BACKGROUND & OBJECTIVES: Polio, measles, rubella, influenza and rotavirus surveillance programmes are of great public health importance globally. Virus isolation using cell culture is an integral part of such programmes. Possibility of unintended isolation of SARS-CoV-2 from clinical specimens processed in biosafety level-2 (BSL-2) laboratories during the above-mentioned surveillance programmes, cannot be ruled out. The present study was conducted to assess the susceptibility of different cell lines to SARS-CoV-2 used in these programmes. METHODS: Replication of SARS-CoV-2 was studied in RD and L20B, Vero/hSLAM, MA-104 and Madin–Darby Canine Kidney (MDCK) cell lines, used for the isolation of polio, measles, rubella, rotavirus and influenza viruses, respectively. SARS-CoV-2 at 0.01 multiplicity of infection was inoculated and the viral growth was assessed by observation of cytopathic effects followed by real-time reverse transcription–polymerase chain reaction (qRT-PCR). Vero CCL-81 cell line was used as a positive control. RESULTS: SARS-CoV-2 replicated in Vero/hSLAM, and MA-104 cells, whereas it did not replicate in L20B, RD and MDCK cells. Vero/hSLAM, and Vero CCL-81 showed rounding, degeneration and detachment of cells; MA-104 cells also showed syncytia formation. In qRT-PCR, Vero/hSLAM and MA-104 showed 10(6) and Vero CCL-81 showed 10(7) viral RNA copies per µl. The 50 per cent tissue culture infectious dose titres of Vero/hSLAM, MA-104 and Vero CCL-81 were 10(5.54), 10(5.29) and 10(6.45)/ml, respectively. INTERPRETATION & CONCLUSIONS: Replication of SARS-CoV-2 in Vero/hSLAM and MA-104 underscores the possibility of its unintended isolation during surveillance procedures aiming to isolate measles, rubella and rotavirus. This could result in accidental exposure to high titres of SARS-CoV-2, which can result in laboratory acquired infections and community risk, highlighting the need for revisiting biosafety measures in public health laboratories.
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spelling pubmed-95523912022-10-12 Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety Pawar, Shailesh D. Kode, Sadhana S. Keng, Sachin S. Tare, Deeksha S. Diop, Ousmane M. Abraham, Priya Sharma, Deepa K. Sangal, Lucky Yadav, Pragya D. Potdar, Varsha A. Indian J Med Res Original Article BACKGROUND & OBJECTIVES: Polio, measles, rubella, influenza and rotavirus surveillance programmes are of great public health importance globally. Virus isolation using cell culture is an integral part of such programmes. Possibility of unintended isolation of SARS-CoV-2 from clinical specimens processed in biosafety level-2 (BSL-2) laboratories during the above-mentioned surveillance programmes, cannot be ruled out. The present study was conducted to assess the susceptibility of different cell lines to SARS-CoV-2 used in these programmes. METHODS: Replication of SARS-CoV-2 was studied in RD and L20B, Vero/hSLAM, MA-104 and Madin–Darby Canine Kidney (MDCK) cell lines, used for the isolation of polio, measles, rubella, rotavirus and influenza viruses, respectively. SARS-CoV-2 at 0.01 multiplicity of infection was inoculated and the viral growth was assessed by observation of cytopathic effects followed by real-time reverse transcription–polymerase chain reaction (qRT-PCR). Vero CCL-81 cell line was used as a positive control. RESULTS: SARS-CoV-2 replicated in Vero/hSLAM, and MA-104 cells, whereas it did not replicate in L20B, RD and MDCK cells. Vero/hSLAM, and Vero CCL-81 showed rounding, degeneration and detachment of cells; MA-104 cells also showed syncytia formation. In qRT-PCR, Vero/hSLAM and MA-104 showed 10(6) and Vero CCL-81 showed 10(7) viral RNA copies per µl. The 50 per cent tissue culture infectious dose titres of Vero/hSLAM, MA-104 and Vero CCL-81 were 10(5.54), 10(5.29) and 10(6.45)/ml, respectively. INTERPRETATION & CONCLUSIONS: Replication of SARS-CoV-2 in Vero/hSLAM and MA-104 underscores the possibility of its unintended isolation during surveillance procedures aiming to isolate measles, rubella and rotavirus. This could result in accidental exposure to high titres of SARS-CoV-2, which can result in laboratory acquired infections and community risk, highlighting the need for revisiting biosafety measures in public health laboratories. Wolters Kluwer - Medknow 2022-01 /pmc/articles/PMC9552391/ /pubmed/35859439 http://dx.doi.org/10.4103/ijmr.ijmr_1448_21 Text en Copyright: © 2022 Indian Journal of Medical Research https://creativecommons.org/licenses/by-nc-sa/4.0/This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.
spellingShingle Original Article
Pawar, Shailesh D.
Kode, Sadhana S.
Keng, Sachin S.
Tare, Deeksha S.
Diop, Ousmane M.
Abraham, Priya
Sharma, Deepa K.
Sangal, Lucky
Yadav, Pragya D.
Potdar, Varsha A.
Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety
title Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety
title_full Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety
title_fullStr Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety
title_full_unstemmed Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety
title_short Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety
title_sort replication of sars-cov-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9552391/
https://www.ncbi.nlm.nih.gov/pubmed/35859439
http://dx.doi.org/10.4103/ijmr.ijmr_1448_21
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