Paper Info
Title
Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection
Abstract
The fast spread of SARS-CoV-2 has led to a global pandemic, calling for fast and accurate assays to allow infection diagnosis and prevention of transmission. We aimed to develop a molecular beacon (MB)-based detection assay for SARS-CoV-2, designed to detect the ORF1ab and S genes, proposing a two-stage COVID-19 testing strategy. The novelty of this work lies in the design and optimization of two MBs for detection of SARS-CoV-2, namely, concentration, fluorescence plateaus of hybridization, reaction temperature and real-time results. We also identify putative G-quadruplex (G4) regions in the genome of SARS-CoV-2. A total of 458 nasopharyngeal and throat swab samples (426 positive and 32 negative) were tested with the MB assay and the fluorescence levels compared with the cycle threshold (Ct) values obtained from a commercial RT-PCR test in terms of test duration, sensitivity, and specificity. Our results show that the samples with higher fluorescence levels correspond to those with low Ct values, suggesting a correlation between viral load and increased MB fluorescence. The proposed assay represents a fast (total duration of 2 h 20 min including amplification and fluorescence reading stages) and simple way of detecting SARS-CoV-2 in clinical samples from the upper respiratory tract.
Year
DOI
Venue
2021
10.3390/s21217015
SENSORS
Keywords
DocType
Volume
RNA viruses, SARS-CoV-2, COVID-19, fluorescence detection, diagnosis
Journal
21
Issue
ISSN
Citations 
21
1424-8220
0
PageRank 
References 
Authors
0.34
0
14
Name
Order
Citations
PageRank
Josué Carvalho100.34
Jéssica Lopes-Nunes200.34
Joana Figueiredo300.34
Tiago Santos411.06
André Miranda500.34
Micaela Riscado600.34
Fani Sousa700.34
Ana Paula Duarte800.34
Sílvia Socorro900.34
Cândida Teixeira Tomaz1000.34
Mafalda Felgueiras1100.34
Rui Teixeira1200.34
Conceição Faria1300.34
Carla Cruz1411.41