Loop-Mediated Isothermal Amplification (LAMP): An Innovative Approach for the Environmental Monitoring of SARS-CoV-2 DOI Creative Commons
Simona Spiteri, Federica Marino, Luna Girolamini

et al.

Pathogens, Journal Year: 2024, Volume and Issue: 13(11), P. 1022 - 1022

Published: Nov. 20, 2024

The rapid and accurate detection of SARS-CoV-2 in environmental settings is crucial for effective public health management during the COVID-19 pandemic. This study compares performance Reverse Transcription quantitative polymerase chain reaction (RT-qPCR) loop-mediated isothermal amplification (RT-LAMP) from 100 surface samples collected healthcare environments. reference method, RT-qPCR, identified a percentage 25% positive samples, while RT-LAMP detected 27% surfaces. Our findings reveal sensitivity 32% specificity 75% RT-LAMP, with predictive value 30% negative 77%. overall accuracy concordance RT-qPCR was 64% both methods. Despite its lower compared to had an advantage due screening surveillance, which particularly useful confirming results. These results underscore potential not only as valuable method monitoring but also system control sanitation process ordinary emergency conditions, providing further optimization validation reliability routine surveillance outbreak response efforts.

Language: Английский

Comprehensive genomic analysis of the SARS-CoV-2 Omicron variant BA.2.76 in Jining City, China, 2022 DOI Creative Commons
Qiang Yin, Wei Liu,

Yajuan Jiang

et al.

BMC Genomics, Journal Year: 2024, Volume and Issue: 25(1)

Published: April 17, 2024

Abstract Objective This study aims to analyze the molecular characteristics of novel coronavirus (SARS-CoV-2) Omicron variant BA.2.76 in Jining City, China. Methods Whole-genome sequencing was performed on 87 cases SARS-CoV-2 infection. Evolutionary trees were constructed using bioinformatics software sequence homology, sites, N-glycosylation and phosphorylation sites. Results All whole-genome sequences classified under evolutionary branch BA.2.76. Their similarity reference strain Wuhan-Hu-1 ranged from 99.72 99.74%. In comparison Wuhan-Hu-1, exhibited 77–84 nucleotide differences 27 deletions. A total 69 amino acid 9 deletions, 1 stop codon mutation identified across 18 proteins. Among them, spike (S) protein highest number ORF8 showed a Q27 mutation. Multiple proteins displayed variations glycosylation Conclusion continues evolve, giving rise new strains with enhanced transmission, stronger immune evasion capabilities, reduced pathogenicity. The application high-throughput technologies epidemic prevention control COVID-19 provides crucial insights into virus at genomic level, thereby holding significant implications for pandemic.

Language: Английский

Citations

4
Revisiting the dimensions of universal vaccine with special focus on COVID-19: Efficacy versus methods of designing DOI

Puja Jaishwal,

Kisalay Jha,

Satarudra Prakash Singh

et al.

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 277, P. 134012 - 134012

Published: July 22, 2024

Language: Английский

Citations

4
Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid Protein DOI Creative Commons
Robert Hnasko,

Alice V. Lin,

Jeffery A. McGarvey

et al.

Microbial Biotechnology, Journal Year: 2025, Volume and Issue: 18(2)

Published: Feb. 1, 2025

ABSTRACT Immunoassays represent sensitive, easy‐to‐use, and cost‐effective tests useful for the detection of SARS‐CoV‐2 virus. In this manuscript, we report on binding specificity a pair novel monoclonal antibodies (MAbs) generated against nucleocapsid protein (NP) their development into sensitive sandwich enzyme‐linked immunosorbent assays (sELISA) lateral flow immunoassay (LFIA). Binding these MAbs to hCoVs is limited variants SARS‐CoV NP. Chemiluminescent absorbance spectroscopy sELISAs limit (LOD) B.1.1.529 NP variant at 15 pg/mL, LFIA using red‐dyed 200 nm particle 10 ng/mL. The sELISA exhibits broad viral with assay LOD virus 1.4 × 5 genome copies per mL ( p ≤ 0.001). availability should facilitate continued investment in commercial immunoassays increase global technologies.

Language: Английский

Citations

0
Immunological assessment of NSFu1: A novel fusion molecule constructed from structural proteins of SARS-CoV-2 for improving COVID-19 antibody detection DOI

Shaista Arif,

Mohsina Akhter,

Aqsa Anwar

et al.

Archives of Microbiology, Journal Year: 2025, Volume and Issue: 207(4)

Published: March 15, 2025

Language: Английский

Citations

0
Measuring SARS-CoV-2 RNA in Bangkok wastewater treatment plants and estimating infected population after fully opening the country in 2023, Thailand DOI Creative Commons

Thanchira Saita,

Bussarakam Thitanuwat,

Nattamon Niyomdecha

et al.

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: March 20, 2025

Wastewater-based epidemiology (WBE) has been employed for monitoring the presence of SARS-CoV-2 infected population. Herein, study aims to apply WBE surveillance and in Bangkok, where highest official covid-19 cases reported Thailand, during fully opening international tourists early 2023. A total 200 wastewater samples (100 influent 100 effluent samples) were collected from 10 treatment plants (WWTPs) January–May RNA was detected by real time qRT-PCR with accounting 51% (102/200). Of these, 88% (88/100) 14% (14/100) samples, respectively. The concentration ranged 4.76 × 102–1.48 105 copies/L. amount increased approximately 4 times lag phase (January–March) log (April–May). Spearman's correlation coefficient revealed that between estimated population weekly statistically significant (p-value = 0.017). had a relationship (r 0.481, p-value < 0.001). Lag analysis warning 1–3 weeks before rising observed. GIS applied spatial-temporal at province level, suggesting dashboard should be further developed.

Language: Английский

Citations

0
Wide Real-Life Data Support Reduced Sensitivity of Antigen Tests for Omicron SARS-CoV-2 Infections DOI Creative Commons
Chiara Piubelli, Davide Treggiari, Denise Lavezzari

et al.

Viruses, Journal Year: 2024, Volume and Issue: 16(5), P. 657 - 657

Published: April 23, 2024

With the continuous spread of new SARS-CoV-2 variants concern (VOCs), monitoring diagnostic test performances is mandatory. We evaluated changes in antigen tests’ (ADTs) accuracy along Delta to Omicron VOCs transition, exploring N protein mutations possibly affecting ADT sensitivity and assessing best sampling site for diagnosis infections. In total, 5175 subjects were enrolled from 1 October 2021 15 July 2022. The inclusion criteria combined with a same-day RT-PCR swab test. For analysis, 61 patients prospectively recruited during period nasal oral analyses by RT-PCR. Next-Generation Sequencing data obtained evaluate different sublineages. Using as reference, 387 resulted becoming infected overall decreased 63% 33% period. This decrease was highly statistically significant (p < 0.001), no viral load detected at RNA level. presented significantly higher than wave. reduced detection rate infections should be considered global testing strategy preserve accurate diagnoses across changing variants.

Language: Английский

Citations

3
Development of Detection Antibody Targeting the Linear Epitope in SARS-CoV-2 Nucleocapsid Protein with Ultra-High Sensitivity DOI Open Access
Feng Wu, Yike Jiang, Hongtian Yang

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(8), P. 4436 - 4436

Published: April 18, 2024

The COVID-19 pandemic caused by SARS-CoV-2 highlighted the importance of reliable detection methods for disease control and surveillance. Optimizing antibodies rational screening antigens would improve sensitivity specificity antibody-based such as colloidal gold immunochromatography. In this study, we screened three peptide with conserved sequences in N protein using bioinformatical structural biological analyses. Antibodies that specifically recognize these peptides were prepared. epitope had highest binding affinity its antibody was located on surface protein, which favorable binding. Using optimal can epitope, developed immunochromatography, detect at 10 pg/mL. Importantly, could effectively both natural antigen mutated showing feasibility being applied large-scale population testing SARS-CoV-2. Our study provides a platform reference significance high sensitivity, specificity, reliability other pathogens.

Language: Английский

Citations

2
Pharmacokinetic and Environmental Risk Assessment of Prime-2-CoV, a Non-Replicating Orf Virus-Based Vaccine against SARS-CoV-2 DOI Creative Commons

C.A. Metz,

Verena Haug, Melanie Müller

et al.

Vaccines, Journal Year: 2024, Volume and Issue: 12(5), P. 492 - 492

Published: May 2, 2024

Viral vector vaccines represent a substantial advancement in immunization technology, offering numerous benefits over traditional vaccine modalities. The Orf virus (ORFV) strain D1701-VrV is particularly promising candidate for development due to its distinctive attributes, such as good safety profile, the ability elicit both humoral and cellular immunity, favorable genetic thermal stability. Despite ORFV's theoretical advantages, narrow host range limited systemic spread post-inoculation, critical gap persists between these empirical evidence regarding vivo profile. This discrepancy underscores need comprehensive preclinical validations bridge this knowledge gap, especially considering use humans. Our research introduces Prime-2-CoV, an innovative ORFV-based against COVID-19, designed robust immune response by expressing SARS-CoV-2 Nucleocapsid Spike proteins. Currently under clinical trials, Prime-2-CoV marks inaugural application of ORFV human subjects. Addressing aforementioned concerns, our extensive evaluation, including environmental risk assessment (ERA) detailed pharmacokinetic studies rats immunocompromised NOG mice, demonstrates Prime-2-CoV's negligible impact, minimal ERA risks. These findings not only affirm vaccine's efficacy but also pioneer therapeutics, highlighting potential wider therapeutic applications.

Language: Английский

Citations

2
Single intranasal immunization with a high dose of influenza vector protects against infection with heterologous influenza virus and SARS-CoV-2 in ferrets and hamsters DOI Creative Commons
Andrej Egorov, Artem Krokhin, И. А. Ленева

et al.

Microbiology Independent Research Journal (MIR Journal), Journal Year: 2024, Volume and Issue: 11(1)

Published: Jan. 1, 2024

BACKGROUND: The challenge of vaccine effectiveness against viruses that undergo constant antigenic changes during evolution is currently being addressed by updating formulations to match circulating strains. However, this approach proves ineffective if a virus undergoes drift and shift, or new virus, such as SARS-CoV-2, emerges enters circulation. Hence, there pressing need develop universal vaccines elicit T-cell immune response targeting conserved determinants pathogens. OBJECTIVE: To candidate influenza coronavirus based on an attenuated vector. METHODS: In pursuit objective, we developed recombinant vector named FluCoV-N. It incorporates attenuating modifications in the ns1 nep genes expresses N-terminal half N protein (N 1-209 ) SARS-CoV-2 virus. assess vector’s protective efficacy influenza, ferrets were infected with heterologous A/Austria/1516645/2022 (H3N2) 25th day after single immunization 9.4 log 10 EID 50 studied test protection coronavirus, hamsters immunized once at dose 8.2 challenged 21 days later. RESULTS: As result NS genomic segment, constructed acquired temperature-sensitive (ts) phenotype demonstrated heightened ability induce type 1 interferons. was harmless animals when administered intranasally high doses exceeding 8.0 . ferrets, intranasal FluCoV-N accelerated resolution infection caused H3N2 Similar led 10,000-fold reduction viral titers lungs second reduced pathology animals. CONCLUSION: A protected from hamsters.

Language: Английский

Citations

2
Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 Virus DOI Creative Commons
Lara González‐Cabaleiro, Carlos Fernández‐Lodeiro, Lorena Vázquez‐Iglesias

et al.

Small Science, Journal Year: 2024, Volume and Issue: 4(11)

Published: Aug. 10, 2024

Lateral flow immunoassays (LFIAs) are easy-to-use antigen tests that provide different signal readouts, with colorimetric readouts being the most commonly used. However, these analytical devices have relatively low sensitivity and produce semiquantitative results, limiting their diagnostic applications. Herein, we address challenges by implementing a digital surface-enhanced Raman spectroscopy (SERS)-based LFIA for accurate ultrasensitive quantitative detection of SARS-CoV-2 nucleocapsid (N) protein. Compared average SERS intensity measurements, approach allowed to overcome fluctuations in scattering signals, thereby increasing assay. Our method exhibited quantification range viral protein nasal swabs from 0.001 10 pg mL-1, limit down 1.9 aM (0.9 fg mL-1), improving LFIAs conventional-SERS-based several orders magnitude. Importantly, this shows an 0.03 TCID50 which is greater than reported other immunoassays. In conclusion, successfully demonstrate robust SARS-CoV-2N at ultralow concentrations. The improvement may pave way translate technology into arena microbes disease biomarkers.

Language: Английский

Citations

2