tetano
Editor, Senior Moderator
Infect Genet Evol
. 2022 Mar 30;105278.
doi: 10.1016/j.meegid.2022.105278. Online ahead of print.
Identification, propagation and molecular characterization of SARS-CoV-2 delta variant isolated from Egyptian COVID-19 patients
Badriyah Alotaibi[SUP] 1 [/SUP], Thanaa A El-Masry[SUP] 2 [/SUP], Mohamed G Seadawy[SUP] 3 [/SUP], Bassem E El-Harty[SUP] 3 [/SUP], Asmaa Saleh[SUP] 4 [/SUP], Ahmed F Gad[SUP] 3 [/SUP], Mostafa F El-Hosseny[SUP] 3 [/SUP], Yasmen F Mahran[SUP] 5 [/SUP], Maisra M El-Bouseary[SUP] 6 [/SUP]
Affiliations
Abstract
The recently emerging coronavirus, severe acute respiratory syndrome coronavirus 2, (SARS-CoV-2) is the causative agent of the Coronavirus disease 2019 (COVID-19) pandemic. Since its discovery in the city of Wahan, China, SARS-CoV-2 has spread rapidly to invade all countries. In addition to its rapid transmission rate, it is characterized by high genetic mutation rates. The aim of this study is to provide an effective method for the isolation and propagation of SARS-CoV-2 in cell lines without any induction of genetic variations. In this study, we isolated SARS-CoV-2 from oro-nasopharyngeal swabs collected from Egyptian patients who were clinically diagnosed with COVID-19. Molecular identification of SARS-CoV-2 was performed by Real-Time Quantitative Reverse Transcription PCR (RT-qPCR). The isolated virus was propagated on Vero E6 cells without applying serial viral passages to avoid any variation of the viral genome. The replication and propagation were confirmed by the results of both RT-qPCR and the cytopathic effect (CPE). Moreover, SARS-CoV-2 was completely inactivated chemically using beta-propiolactone (βPL). Whole genome sequencing (WGS) of the propagated virus was performed in order to investigate mutational patterns. The genome sequences recovered in 2020 (n = 18) were similar to the reference strain, Wuhan-Hu-1, and were clustered as clade 20A. However, the genomic sequences recovered in 2021 (n = 2) were clustered as clade 21 J. These two sequences are considered the first Delta (B.1.617.2) variants detected in Egypt. This study provides a reference for researchers in Egypt to isolate and propagate SARS-CoV-2 easily and efficiently. Furthermore, the prevalence of the SARS-CoV-2 delta variant in Egypt necessitates continuous monitoring of the efficacy of the applied treatment protocol and the effectiveness of current vaccines against such variants of concern (VOC).
Keywords: Delta variant; Isolation; Propagation; RT-qPCR; SARS-CoV-2; Variant of concern.
. 2022 Mar 30;105278.
doi: 10.1016/j.meegid.2022.105278. Online ahead of print.
Identification, propagation and molecular characterization of SARS-CoV-2 delta variant isolated from Egyptian COVID-19 patients
Badriyah Alotaibi[SUP] 1 [/SUP], Thanaa A El-Masry[SUP] 2 [/SUP], Mohamed G Seadawy[SUP] 3 [/SUP], Bassem E El-Harty[SUP] 3 [/SUP], Asmaa Saleh[SUP] 4 [/SUP], Ahmed F Gad[SUP] 3 [/SUP], Mostafa F El-Hosseny[SUP] 3 [/SUP], Yasmen F Mahran[SUP] 5 [/SUP], Maisra M El-Bouseary[SUP] 6 [/SUP]
Affiliations
- PMID: 35367360
- DOI: 10.1016/j.meegid.2022.105278
Abstract
The recently emerging coronavirus, severe acute respiratory syndrome coronavirus 2, (SARS-CoV-2) is the causative agent of the Coronavirus disease 2019 (COVID-19) pandemic. Since its discovery in the city of Wahan, China, SARS-CoV-2 has spread rapidly to invade all countries. In addition to its rapid transmission rate, it is characterized by high genetic mutation rates. The aim of this study is to provide an effective method for the isolation and propagation of SARS-CoV-2 in cell lines without any induction of genetic variations. In this study, we isolated SARS-CoV-2 from oro-nasopharyngeal swabs collected from Egyptian patients who were clinically diagnosed with COVID-19. Molecular identification of SARS-CoV-2 was performed by Real-Time Quantitative Reverse Transcription PCR (RT-qPCR). The isolated virus was propagated on Vero E6 cells without applying serial viral passages to avoid any variation of the viral genome. The replication and propagation were confirmed by the results of both RT-qPCR and the cytopathic effect (CPE). Moreover, SARS-CoV-2 was completely inactivated chemically using beta-propiolactone (βPL). Whole genome sequencing (WGS) of the propagated virus was performed in order to investigate mutational patterns. The genome sequences recovered in 2020 (n = 18) were similar to the reference strain, Wuhan-Hu-1, and were clustered as clade 20A. However, the genomic sequences recovered in 2021 (n = 2) were clustered as clade 21 J. These two sequences are considered the first Delta (B.1.617.2) variants detected in Egypt. This study provides a reference for researchers in Egypt to isolate and propagate SARS-CoV-2 easily and efficiently. Furthermore, the prevalence of the SARS-CoV-2 delta variant in Egypt necessitates continuous monitoring of the efficacy of the applied treatment protocol and the effectiveness of current vaccines against such variants of concern (VOC).
Keywords: Delta variant; Isolation; Propagation; RT-qPCR; SARS-CoV-2; Variant of concern.