Molecular beacon assay development for Severe Acute Respiratory Syndrome Coronavirus 2 detection

• DOI: 10.22541/au.161652048.85831942/v1
• Publication Date: 2021-03-23T17:28:25Z
• Authors: Josué Carvalho; Jéssica Lopes Nunes; Joana Figueiredo; Tiago Santos; André Miranda; Micaela Riscado; Fani Sousa; Ana Paula Duarte; Silvia Socorro; Cândida Tomaz; Mafalda Felgueiras; Rui Teixeira; Conceição Faria; Carla Cruz
• Keywords: Amplicon; Molecular beacon

The fast spread of SARS-CoV-2 has led to a global pandemic, calling for and accurate assays allow infection diagnosis prevention transmission. We aimed develop molecular beacon (MB)-based detection assay SARS-CoV-2, designed to, detect the ORF1ab S genes, proposing two-stage COVID-19 testing strategy, using MBs presence target amplicons by fluorescence analysis. Two were designed, optimized in terms concentration, plateaus hybridization, reaction temperature best real-time results. A total 450 nasopharyngel throat swab samples (418 positive 32 negative) tested with MB levels compared cycle threshold (Ct) values obtained from commercial RT-PCR test duration, sensitivity specificity. Our results show that higher correspond those low Ct values, suggesting correlation between viral load increased fluorescence. proposed represents (total duration 2 h 20 min including amplification reading stages) simple way detecting clinical upper respiratory tract. strategy is suitable further development into point-of-care potentially scalable population level.