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...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Medknow Publications
2022-01-01
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| Series: | Indian Journal of Medical Research |
| Subjects: | |
| Online Access: | http://www.ijmr.org.in/article.asp?issn=0971-5916;year=2022;volume=155;issue=1;spage=129;epage=135;aulast=Pawar |
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| author | Shailesh D Pawar Sadhana S Kode Sachin S Keng Deeksha S Tare Ousmane M Diop Priya Abraham Deepa K Sharma Lucky Sangal Pragya D Yadav Varsha A Potdar |
| author_facet | Shailesh D Pawar Sadhana S Kode Sachin S Keng Deeksha S Tare Ousmane M Diop Priya Abraham Deepa K Sharma Lucky Sangal Pragya D Yadav Varsha A Potdar |
| author_sort | Shailesh D Pawar |
| collection | DOAJ |
| 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 106 and Vero CCL-81 showed 107 viral RNA copies per μl. The 50 per cent tissue culture infectious dose titres of Vero/hSLAM, MA-104 and Vero CCL-81 were 105.54, 105.29 and 106.45/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. |
| first_indexed | 2024-04-11T21:41:12Z |
| format | Article |
| id | doaj.art-819fc5d3de074081b3a30cf8b96c5333 |
| institution | Directory Open Access Journal |
| issn | 0971-5916 |
| language | English |
| last_indexed | 2024-04-11T21:41:12Z |
| publishDate | 2022-01-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Indian Journal of Medical Research |
| spelling | doaj.art-819fc5d3de074081b3a30cf8b96c53332022-12-22T04:01:35ZengWolters Kluwer Medknow PublicationsIndian Journal of Medical Research0971-59162022-01-01155112913510.4103/ijmr.ijmr_1448_21Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafetyShailesh D PawarSadhana S KodeSachin S KengDeeksha S TareOusmane M DiopPriya AbrahamDeepa K SharmaLucky SangalPragya D YadavVarsha A PotdarBackground & 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 106 and Vero CCL-81 showed 107 viral RNA copies per μl. The 50 per cent tissue culture infectious dose titres of Vero/hSLAM, MA-104 and Vero CCL-81 were 105.54, 105.29 and 106.45/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.http://www.ijmr.org.in/article.asp?issn=0971-5916;year=2022;volume=155;issue=1;spage=129;epage=135;aulast=Pawarbiosafety - cell lines - covid-19 - sars-cov-2 - surveillance - virus replication |
| spellingShingle | Shailesh D Pawar Sadhana S Kode Sachin S Keng Deeksha S Tare Ousmane M Diop Priya Abraham Deepa K Sharma Lucky Sangal Pragya D Yadav Varsha A Potdar Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety Indian Journal of Medical Research biosafety - cell lines - covid-19 - sars-cov-2 - surveillance - virus replication |
| 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 | biosafety - cell lines - covid-19 - sars-cov-2 - surveillance - virus replication |
| url | http://www.ijmr.org.in/article.asp?issn=0971-5916;year=2022;volume=155;issue=1;spage=129;epage=135;aulast=Pawar |
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