Detecting SARS-CoV-2 cryptic lineages using publicly available whole genome wastewater sequencing data.

Beginning in early 2021, unique and highly divergent lineages of SARS-CoV-2 were sporadically found in wastewater sewersheds using a sequencing strategy focused on amplifying the most rapidly evolving region of SARS-CoV-2, the receptor binding domain (RBD). Because these RBD sequences did not match...

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Main Authors: Reinier Suarez, Devon A Gregory, David A Baker, Clayton A Rushford, Torin L Hunter, Nicholas R Minor, Clayton M Russ, Emma E Copen, David H O'Connor, Marc C Johnson
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-06-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1012850
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author Reinier Suarez
Devon A Gregory
David A Baker
Clayton A Rushford
Torin L Hunter
Nicholas R Minor
Clayton M Russ
Emma E Copen
David H O'Connor
Marc C Johnson
author_facet Reinier Suarez
Devon A Gregory
David A Baker
Clayton A Rushford
Torin L Hunter
Nicholas R Minor
Clayton M Russ
Emma E Copen
David H O'Connor
Marc C Johnson
author_sort Reinier Suarez
collection DOAJ
description Beginning in early 2021, unique and highly divergent lineages of SARS-CoV-2 were sporadically found in wastewater sewersheds using a sequencing strategy focused on amplifying the most rapidly evolving region of SARS-CoV-2, the receptor binding domain (RBD). Because these RBD sequences did not match known circulating strains and their source was not known, we termed them "cryptic lineages". To date, more than 20 cryptic lineages have been identified using the RBD-focused sequencing strategy. Here, we identified and characterized additional cryptic lineages from SARS-CoV-2 wastewater sequences submitted to NCBI's Sequence Read Archives (SRA). Wastewater sequence datasets were screened for individual sequence reads that contained combinations of mutations frequently found in cryptic lineages but not contemporary circulating lineages. Using this method, we identified 18 cryptic lineages that appeared in multiple (2-81) samples from the same sewershed, including 12 that were not previously reported. Partial consensus sequences were generated for each cryptic lineage by extracting and mapping sequences containing cryptic-specific mutations. Surprisingly, seven of the mutations that appeared convergently in cryptic lineages were reversions to sequences that were highly conserved in SARS-CoV-2-related enteric bat Sarbecoviruses. The apparent reversion to bat Sarbecovirus sequences is consistent with the notion that SARS-CoV-2 adaptation to replicate efficiently in respiratory tissues preceded the COVID-19 pandemic.
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issn 1553-7366
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spelling doaj-art-111244d9ce284f0083bb09bf1232027a2025-08-20T02:36:53ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742025-06-01216e101285010.1371/journal.ppat.1012850Detecting SARS-CoV-2 cryptic lineages using publicly available whole genome wastewater sequencing data.Reinier SuarezDevon A GregoryDavid A BakerClayton A RushfordTorin L HunterNicholas R MinorClayton M RussEmma E CopenDavid H O'ConnorMarc C JohnsonBeginning in early 2021, unique and highly divergent lineages of SARS-CoV-2 were sporadically found in wastewater sewersheds using a sequencing strategy focused on amplifying the most rapidly evolving region of SARS-CoV-2, the receptor binding domain (RBD). Because these RBD sequences did not match known circulating strains and their source was not known, we termed them "cryptic lineages". To date, more than 20 cryptic lineages have been identified using the RBD-focused sequencing strategy. Here, we identified and characterized additional cryptic lineages from SARS-CoV-2 wastewater sequences submitted to NCBI's Sequence Read Archives (SRA). Wastewater sequence datasets were screened for individual sequence reads that contained combinations of mutations frequently found in cryptic lineages but not contemporary circulating lineages. Using this method, we identified 18 cryptic lineages that appeared in multiple (2-81) samples from the same sewershed, including 12 that were not previously reported. Partial consensus sequences were generated for each cryptic lineage by extracting and mapping sequences containing cryptic-specific mutations. Surprisingly, seven of the mutations that appeared convergently in cryptic lineages were reversions to sequences that were highly conserved in SARS-CoV-2-related enteric bat Sarbecoviruses. The apparent reversion to bat Sarbecovirus sequences is consistent with the notion that SARS-CoV-2 adaptation to replicate efficiently in respiratory tissues preceded the COVID-19 pandemic.https://doi.org/10.1371/journal.ppat.1012850
spellingShingle Reinier Suarez
Devon A Gregory
David A Baker
Clayton A Rushford
Torin L Hunter
Nicholas R Minor
Clayton M Russ
Emma E Copen
David H O'Connor
Marc C Johnson
Detecting SARS-CoV-2 cryptic lineages using publicly available whole genome wastewater sequencing data.
PLoS Pathogens
title Detecting SARS-CoV-2 cryptic lineages using publicly available whole genome wastewater sequencing data.
title_full Detecting SARS-CoV-2 cryptic lineages using publicly available whole genome wastewater sequencing data.
title_fullStr Detecting SARS-CoV-2 cryptic lineages using publicly available whole genome wastewater sequencing data.
title_full_unstemmed Detecting SARS-CoV-2 cryptic lineages using publicly available whole genome wastewater sequencing data.
title_short Detecting SARS-CoV-2 cryptic lineages using publicly available whole genome wastewater sequencing data.
title_sort detecting sars cov 2 cryptic lineages using publicly available whole genome wastewater sequencing data
url https://doi.org/10.1371/journal.ppat.1012850
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