Integration of Whole-Genome Sequencing with ddPCR Kit for Detection of Omicron Subvariants in Wastewater in the Upper Peninsula of Michigan DOI Creative Commons
Michelle M. Jarvie,

Thu N.T. Nguyen,

Benjamin Southwell

и другие.

Applied Microbiology, Год журнала: 2024, Номер 4(4), С. 1453 - 1463

Опубликована: Окт. 13, 2024

This study explores the integration of genome sequencing and digital droplet polymerase chain reaction (ddPCR)-based methods for tracking diversity COVID-19 variants in wastewater. The research focuses on monitoring various Omicron subvariants during a period significant viral evolution. Genome sequencing, particularly using Oxford Nanopore Technology (ONT), provides detailed view emerging variants, surpassing limitations PCR-based detection kits that rely known sequences. Of 43 samples analyzed, 39.5% showed matching results between GT Molecular ddPCR though only 4% were exact matches. Some mismatches occurred due to newer like XBB BQ.1, which could not detect. emphasized kits, variant sequences, while real-time data offering more comprehensive circulating strains. highlights effectiveness combining these methodologies enhance early inform public health strategies, especially regions with limited clinical capabilities.

Язык: Английский

Long-Term Immune Consequences of Initial SARS-CoV-2 A.23.1 Exposure: A Longitudinal Study of Antibody Responses and Cross-Neutralization in a Ugandan Cohort DOI Creative Commons

Gerald Kevin Oluka,

Jackson Sembera,

Joseph Ssebwana Katende

и другие.

Vaccines, Год журнала: 2025, Номер 13(2), С. 143 - 143

Опубликована: Янв. 29, 2025

Background: This study assessed the long-term dynamics of neutralizing antibodies in a Ugandan cohort primarily exposed to A.23.1 SARS-CoV-2 variant, examining how this shaped immune breadth and potency against diverse strains following infection prototype-based vaccination. Methods: We conducted 427-day retrospective analysis 41 participants across multiple waves, assessing binding antibody responses using in-house ELISA pseudotyped virus neutralization assays. quantified key variants, A.23.1, D614G, Delta, BA.4, capturing evolving immunity pandemic. Results: Neutralizing titers remained significantly higher than those highlighting solid memory infection. Consistently lower were observed for BA.4 all time points, aligning with its strong immune-evasion capability. Correlations between spike-directed IgG (S-IgG) concentrations stronger no correlation BA.4. ChAdOx1-S vaccination substantially elevated most notably essential role boosting immunity, even individuals initially low titers. Conclusions: Initial exposure variant triggered potent responses, shaping during subsequent exposures. These findings highlight importance accounting early viral exposures vaccine development public health planning. The distinctly response highlights need continuous antigenic monitoring timely updates protection emerging variants. Vaccination remains reinforcing sustaining

Язык: Английский

Процитировано

1

Adaptive evolution of SARS-CoV-2 during a persistent infection for 521 days in an immunocompromised patient DOI Creative Commons
Hanno Schmidt, Luca Aroha Schick, Jürgen Podlech

и другие.

npj Genomic Medicine, Год журнала: 2025, Номер 10(1)

Опубликована: Янв. 17, 2025

Immunocompromised patients struggle to adequately clear viral infections, offering the virus opportunity adapt immune system in host. Here we present a case study of patient undergoing allogeneic hematopoietic stem cell transplantation with 521-day follow-up SARS-CoV-2 infection BF.7.21 variant. Virus samples from five time points were submitted whole genome sequencing. Between first detection and its clearance, patient's population acquired 34 amino acid substitutions 8 deletions coding regions. With 11 receptor binding domain virus' spike protein, 15 times more abundant than expected for random distribution this highly functional region. Amongst them S:K417T, S:N440S, S:K444R, S:V445A, S:G446N, S:L452Q, S:N460K, S:E484V at positions that are notorious their resistance-mediating effects. The substitution patterns found indicate ongoing adaptive evolution.

Язык: Английский

Процитировано

0

Role of N-linked glycosylation sites in human ACE2 in SARS-CoV-2 and hCoV-NL63 infection DOI Creative Commons
Sabrina Noettger, Fabian Zech, Rayhane Nchioua

и другие.

Journal of Virology, Год журнала: 2025, Номер unknown

Опубликована: Март 28, 2025

ABSTRACT Angiotensin-converting enzyme 2 (ACE2) is a transmembrane protein known for its physiological role in the renin-angiotensin system that also serves as receptor entry of SARS-CoV-1, SARS-CoV-2, and seasonal human coronavirus NL63 (hCoV-NL63). ACE2 contains seven N-linked glycosylation sites. Molecular simulation binding analyses suggest some them are involved interaction with Spike (S) proteins hCoVs, but their relevance S-mediated fusion viral poorly investigated. To address this, we determined impact all sites on SARS-CoV-2 hCoV-NL63 infection well cell-to-cell fusion. We found mutant expressed localized at cell surface, albeit lacks glycans decreased levels. On average, changes T92I, N322A, N690A, combined mutation increased endocytic VSVpp mediated by early HU-1 Omicron BA.2, BA.5, XBB.1.5 S proteins. In comparison, only lack glycan N322 enhanced syncytia formation case Changes N90A, N322A strain about twofold to threefold had lesser effects genuine variants. Despite reduced surface expression ACE2, elimination usually via pathway while having little effect presence TMPRSS2. Our results provide insights into ability (hACE2) serve receptors infection. IMPORTANCE Several coronaviruses use angiotensin-converting primary cells. glycosylated distinct positions, target cells incompletely understood. Here, examined individual mutations hACE2 Spike-mediated VSV-pseudoparticle new information highly pathogenic coronaviruses.

Язык: Английский

Процитировано

0

CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex DOI Creative Commons
Ildar Gabaev,

Alexandra Rowland,

Emilija Jovanovic

и другие.

Scientific Reports, Год журнала: 2025, Номер 15(1)

Опубликована: Апрель 7, 2025

Abstract The TMPRSS2 cell surface protease is used by a broad range of respiratory viruses to facilitate entry into target cells. Together with ACE2, represents key factor for SARS-CoV-2 infection, as mediates cleavage viral spike protein, enabling direct fusion the envelope host membrane. Since start COVID-19 pandemic, has gained attention therapeutic inhibitors which would inhibit but little known about regulation, particularly in types physiologically relevant infection. Here, we performed an unbiased genome-wide CRISPR-Cas9 library screen, together targeted at epigenetic modifiers and transcriptional regulators, identify cellular factors that modulate expression human colon epithelial We find endogenous regulated Elongin BC-VHL complex HIF transcription factors. Depletion B or treatment cells PHD resulted downregulation inhibition show still utilised Omicron variants colonic Our study enhances our understanding regulation

Язык: Английский

Процитировано

0

Causes and Consequences of Coronavirus Spike Protein Variability DOI Open Access
Fabian Zech, Christoph Jung, Timo Jacob

и другие.

Опубликована: Янв. 3, 2024

Coronaviruses are a large family of diverse enveloped RNA viruses found in numerous animal species. They well-known for their ability to cross species barriers and have been transmitted from bats or intermediate hosts humans on several occasions. Four the seven human coronaviruses (hCoVs) responsible approximately 20% common colds (hCoV-229E, -NL63, -OC43, -HKU1). Two others (SARS-CoV-1 MERS-CoV) cause severe frequently lethal respiratory syndromes but only spread very limited extents populations. In contrast, SARS-CoV-2, while exhibiting pathogenicity, has profound impact public health due its enormous spread. this review, we discuss which initial features SARS-CoV-2 Spike protein subsequent adaptations new host may helped pathogen COVID-19 pandemic. Our focus is forces driving changes consequences virus infectivity, immune evasion resistance preventive therapeutic agents. addition, significance perspectives broad-spectrum therapeutics vaccines.

Язык: Английский

Процитировано

1

Attenuated replication and damaging effects of SARS-CoV-2 Omicron variants in an intestinal epithelial barrier model DOI Creative Commons
Meta Volčič, Rayhane Nchioua, Chiara Pastorio

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Фев. 28, 2024

ABSTRACT Many COVID-19 patients suffer from gastrointestinal symptoms and impaired intestinal barrier function may play a key role in Long COVID. Despite its importance, the impact of SARS-CoV-2 on epithelia is poorly understood. To address this, we established an model integrating epithelial Caco-2 cells, mucus-secreting HT29 cells human Raji cells. This gut allows efficient differentiation into microfold-like faithfully mimics function, highly permissive to infection. Early strains Delta variant replicated with high efficiency, severely disrupted depleted tight junction proteins, such as claudin-1, occludin ZO-1. In comparison, Omicron subvariants also ZO-1 junctions but had fewer damaging effects mucosal integrity function. Remdesivir TMPRSS2 inhibitor Camostat prevented replication thus damage, while Cathepsin E64d was ineffective. Our results support that disrupts further suggest circulating variants are less than earlier viral strains.

Язык: Английский

Процитировано

1

Causes and Consequences of Coronavirus Spike Protein Variability DOI Creative Commons
Fabian Zech, Christoph Jung, Timo Jacob

и другие.

Viruses, Год журнала: 2024, Номер 16(2), С. 177 - 177

Опубликована: Янв. 25, 2024

Coronaviruses are a large family of enveloped RNA viruses found in numerous animal species. They well known for their ability to cross species barriers and have been transmitted from bats or intermediate hosts humans on several occasions. Four the seven human coronaviruses (hCoVs) responsible approximately 20% common colds (hCoV-229E, -NL63, -OC43, -HKU1). Two others (SARS-CoV-1 MERS-CoV) cause severe frequently lethal respiratory syndromes but only spread very limited extents population. In contrast most recent hCoV, SARS-CoV-2, while exhibiting pathogenicity, has profound impact public health due its enormous spread. this review, we discuss which initial features SARS-CoV-2 Spike protein subsequent adaptations new host may helped pathogen COVID-19 pandemic. Our focus is forces driving changes consequences virus infectivity, immune evasion resistance preventive therapeutic agents. addition, briefly address significance perspectives broad-spectrum therapeutics vaccines.

Язык: Английский

Процитировано

0

Evaluating the Quality of Studies Assessing COVID-19 Vaccine Neutralizing Antibody Immunogenicity DOI Creative Commons
Maeva Katzmarzyk, Robert Naughton, Ioannis Sitaras

и другие.

Vaccines, Год журнала: 2024, Номер 12(11), С. 1238 - 1238

Опубликована: Окт. 30, 2024

COVID-19 vaccine-neutralizing antibodies provide early data on potential vaccine effectiveness, but their usefulness depends study reliability and reporting quality.

Язык: Английский

Процитировано

0

Integration of Whole-Genome Sequencing with ddPCR Kit for Detection of Omicron Subvariants in Wastewater in the Upper Peninsula of Michigan DOI Creative Commons
Michelle M. Jarvie,

Thu N.T. Nguyen,

Benjamin Southwell

и другие.

Applied Microbiology, Год журнала: 2024, Номер 4(4), С. 1453 - 1463

Опубликована: Окт. 13, 2024

This study explores the integration of genome sequencing and digital droplet polymerase chain reaction (ddPCR)-based methods for tracking diversity COVID-19 variants in wastewater. The research focuses on monitoring various Omicron subvariants during a period significant viral evolution. Genome sequencing, particularly using Oxford Nanopore Technology (ONT), provides detailed view emerging variants, surpassing limitations PCR-based detection kits that rely known sequences. Of 43 samples analyzed, 39.5% showed matching results between GT Molecular ddPCR though only 4% were exact matches. Some mismatches occurred due to newer like XBB BQ.1, which could not detect. emphasized kits, variant sequences, while real-time data offering more comprehensive circulating strains. highlights effectiveness combining these methodologies enhance early inform public health strategies, especially regions with limited clinical capabilities.

Язык: Английский

Процитировано

0