tetano
Editor, Senior Moderator
Sci Total Environ
. 2021 Mar 8;778:146198.
doi: 10.1016/j.scitotenv.2021.146198. Online ahead of print.
The presence of SARS-CoV-2 RNA in human sewage in Santa Catarina, Brazil, November 2019
Gislaine Fongaro[SUP] 1 [/SUP], Patr?cia Hermes Stoco[SUP] 2 [/SUP], Doris Sobral Marques Souza[SUP] 1 [/SUP], Edmundo Carlos Grisard[SUP] 2 [/SUP], Maria Elisa Magri[SUP] 3 [/SUP], Paula Rogovski[SUP] 1 [/SUP], Marcos Andr? Sch?rner[SUP] 4 [/SUP], Fernando Hartmann Barazzetti[SUP] 4 [/SUP], Ana Paula Christoff[SUP] 5 [/SUP], Luiz Felipe Valter de Oliveira[SUP] 6 [/SUP], Maria Luiza Bazzo[SUP] 4 [/SUP], Glauber Wagner[SUP] 7 [/SUP], Marta Hern?ndez[SUP] 8 [/SUP], David Rodr?guez-L?zaro[SUP] 9 [/SUP]
Affiliations
Abstract
Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) from October 2019 until March 2020. Twenty five ml of sewage samples were clarified and viruses concentrated using a glycine buffer method coupled with polyethylene glycol precipitation, and viral RNA extracted using a commercial kit. SARS-CoV-2 RNA was detected by RT-qPCR using oligonucleotides targeting N1, S and two RdRp regions. The results of all positive samples were further confirmed by a different RT-qPCR system in an independent laboratory. S and RdRp amplicons were sequenced to confirm identity with SARS-CoV-2. Genome sequencing was performed using two strategies; a sequence-independent single-primer amplification (SISPA) approach, and by direct metagenomics using Illumina's NGS. SARS-CoV-2 RNA was detected on 27th November 2019 (5.49 ? 0.02 log[SUB]10[/SUB] SARS-CoV-2 genome copies (GC) L[SUP]-1[/SUP]), detection being confirmed by an independent laboratory and genome sequencing analysis. The samples in the subsequent three events were positive by all RT-qPCR assays; these positive results were also confirmed by an independent laboratory. The average load was 5.83 ? 0.12 log[SUB]10[/SUB] SARS-CoV-2 GC L[SUP]-1[/SUP], ranging from 5.49 ? 0.02 log[SUB]10[/SUB] GC L[SUP]-1[/SUP] (27th November 2019) to 6.68 ? 0.02 log[SUB]10[/SUB] GC L[SUP]-1[/SUP] (4th March 2020). Our findings demonstrate that SARS-CoV-2 was likely circulating undetected in the community in Brazil since November 2019, earlier than the first reported case in the Americas (21st January 2020).
Keywords: Brazil; COVID-19; Epidemiology; Human sewage; SARS-CoV-2; Surveillance.
. 2021 Mar 8;778:146198.
doi: 10.1016/j.scitotenv.2021.146198. Online ahead of print.
The presence of SARS-CoV-2 RNA in human sewage in Santa Catarina, Brazil, November 2019
Gislaine Fongaro[SUP] 1 [/SUP], Patr?cia Hermes Stoco[SUP] 2 [/SUP], Doris Sobral Marques Souza[SUP] 1 [/SUP], Edmundo Carlos Grisard[SUP] 2 [/SUP], Maria Elisa Magri[SUP] 3 [/SUP], Paula Rogovski[SUP] 1 [/SUP], Marcos Andr? Sch?rner[SUP] 4 [/SUP], Fernando Hartmann Barazzetti[SUP] 4 [/SUP], Ana Paula Christoff[SUP] 5 [/SUP], Luiz Felipe Valter de Oliveira[SUP] 6 [/SUP], Maria Luiza Bazzo[SUP] 4 [/SUP], Glauber Wagner[SUP] 7 [/SUP], Marta Hern?ndez[SUP] 8 [/SUP], David Rodr?guez-L?zaro[SUP] 9 [/SUP]
Affiliations
- PMID: 33714813
- DOI: 10.1016/j.scitotenv.2021.146198
Abstract
Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) from October 2019 until March 2020. Twenty five ml of sewage samples were clarified and viruses concentrated using a glycine buffer method coupled with polyethylene glycol precipitation, and viral RNA extracted using a commercial kit. SARS-CoV-2 RNA was detected by RT-qPCR using oligonucleotides targeting N1, S and two RdRp regions. The results of all positive samples were further confirmed by a different RT-qPCR system in an independent laboratory. S and RdRp amplicons were sequenced to confirm identity with SARS-CoV-2. Genome sequencing was performed using two strategies; a sequence-independent single-primer amplification (SISPA) approach, and by direct metagenomics using Illumina's NGS. SARS-CoV-2 RNA was detected on 27th November 2019 (5.49 ? 0.02 log[SUB]10[/SUB] SARS-CoV-2 genome copies (GC) L[SUP]-1[/SUP]), detection being confirmed by an independent laboratory and genome sequencing analysis. The samples in the subsequent three events were positive by all RT-qPCR assays; these positive results were also confirmed by an independent laboratory. The average load was 5.83 ? 0.12 log[SUB]10[/SUB] SARS-CoV-2 GC L[SUP]-1[/SUP], ranging from 5.49 ? 0.02 log[SUB]10[/SUB] GC L[SUP]-1[/SUP] (27th November 2019) to 6.68 ? 0.02 log[SUB]10[/SUB] GC L[SUP]-1[/SUP] (4th March 2020). Our findings demonstrate that SARS-CoV-2 was likely circulating undetected in the community in Brazil since November 2019, earlier than the first reported case in the Americas (21st January 2020).
Keywords: Brazil; COVID-19; Epidemiology; Human sewage; SARS-CoV-2; Surveillance.