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Since the beginning of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pandemic, it has been clear that testing large groups of the population was the key to stem infection and prevent the effects of the coronavirus disease of 2019, mostly among sensitive patients. On the other hand, time and cost-sustainability of virus detection by molecular analysis such as reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) may be a major issue if testing is extended to large communities, mainly asymptomatic large communities. In this context, sample-pooling and test grouping could offer an effective solution. Here we report the screening on 1195 oral-nasopharyngeal swabs collected from students and staff of the Università degli Studi del Sannio (University of Sannio, Benevento, Campania, Italy) and analyzed by an in-house developed multiplex RT-qPCR for SARS-CoV-2 detection through a simple monodimensional sample pooling strategy. Overall, 400 distinct pools were generated and, within 24 h after swab collection, five positive samples were identified. Out of them, four were confirmed by using a commercially available kit suitable for in vitro diagnostic use (IVD). High accuracy, sensitivity and specificity were also determined by comparing our results with a reference IVD assay for all deconvoluted samples. Overall, we conducted 463 analyses instead of 1195, reducing testing resources by more than 60% without lengthening diagnosis time and without significant losses in sensitivity, suggesting that our strategy was successful in recognizing positive cases in a community of asymptomatic individuals with minor requirements of reagents and time when compared to normal testing procedures.
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http://dx.doi.org/10.3390/diagnostics11071166 | DOI Listing |
PLOS Glob Public Health
September 2025
Department of Biology, University of Ottawa, Ottawa, Ontario, Canada.
Built environment surveillance has shown promise for monitoring COVID-19 burden at granular geographic scales, but its utility for surveillance across larger areas and populations is unknown. Our study aims to evaluate the role of built environment detection of SARS-CoV-2 for the surveillance of COVID-19 across broad geographies and populations. We conducted a prospective city-wide sampling study to examine the relationship between SARS-CoV-2 on floors and COVID-19 burden.
View Article and Find Full Text PDFJ Infect Dev Ctries
August 2025
Department of Medical Microbiology, Faculty of Medicine, Ege University, Izmir 35100, Turkey.
Introduction: The aim of this study was to compare the performance of different clinical specimens-nasopharyngeal (NP) swabs collected by healthcare professionals (HCP-NP), self-collected nasal swabs (Sc-N), and saliva samples (S)-in diagnostic tests for investigating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA and influenza A/B RNA.
Methodology: These clinical samples were collected from 404 symptomatic cases and tested with the SARS-CoV-2 and influenza A/B RNA tests on the cobas 6800 System of Roche Molecular Systems (Roche Molecular Systems, Pleasanton, USA). The SARS-CoV-2 or influenza virus infection status was determined for all patients based on the predefined criteria and corresponding algorithms.
Microbiol Spectr
September 2025
Laboratoy of Virology, Microbiology Department, Hospital Universitario 12 de Octubre, Madrid, Spain.
Millions of reverse transcription-polymerase chain reaction (RT-PCR) tests have been performed worldwide during the SARS-CoV-2 pandemic, using various protocols. This study evaluates the duration of SARS-CoV-2 RNA detectability by RT-PCR at body temperature and analyzes changes in cycle threshold (Ct) values over time. Positive nasopharyngeal swabs for SARS-CoV-2 RT-PCR ( = 120) with different Ct values were collected from Hospital Universitario 12 de Octubre (Madrid, Spain, 2020).
View Article and Find Full Text PDFMicrobiol Spectr
September 2025
Department of Clinical Microbiology, Hospital Clínic of Barcelona-ISGlobal, University of Barcelona, Barcelona, Spain.
Unlabelled: Accurate methods to assess viral viability are crucial for determining isolation duration and antiviral therapy in immunocompromised patients. Although cell culture (CC) is the gold standard, it has limitations. Cycle threshold (Ct) values from genomic RNA (gRNA) RT-PCR and subgenomic RNA (sgRNA) RT-PCR have been proposed as markers of active viral replication.
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