Abstract
Background
So far, one of the major drawbacks of the available molecular assays for the diagnosis of severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) is the need for viral nucleic acid extraction from clinical specimens.
Objective
The aim of this study was to evaluate the performances of a newly designed real-time RT-PCR (Simplexa™ COVID-19 Direct assay), that is established with an all-in-one reagent mix and no separate extraction required.
Results
The lower limit of detection (LOD) for both target genes resulted the same: 3.2 (CI: 2.9–3.8) log10 cp/mL and 0.40 (CI: 0.2–1.5) TCID50/mL for S gene while 3.2 log10 (CI: 2.9–3.7) log10 cp/mL and 0.4 (CI: 0.2–1.3) TCID50/mL for ORF1ab. The LOD obtained with extracted viral RNA for both S gene or ORF1ab was 2.7 log10 cp/mL. Crossreactive analysis performed in 20 nasopharyngeal swabs confirmed a 100% of clinical specificity of the assay. Clinical performances of Simplexa™ COVID-19 Direct assay were assessed in 278 nasopharyngeal swabs tested in parallel with Corman’s method. Concordance analysis showed an “almost perfect” agreement in SARS-CoV-2 RNA detection between the two assays, being κ = 0.938; SE = 0.021; 95% CI = 0.896-0.980.
Conclusions
The high sensitivity and specificity of this new assay indicate that it is promising for laboratory diagnosis, enabling highspeed detection in just over one hour, which is significantly faster than the up to five hours currently required by traditional extraction followed by amplification technologies, thus allowing prompt decision making regarding isolation of infected patients.
【저자키워드】 SARS-CoV-2, Surveillance, real-time RT-PCR, rapid response, 【초록키워드】 COVID-19, Diagnosis, Laboratory, Nasopharyngeal swab, amplification, Sensitivity and specificity, clinical, limit of detection, Isolation, Viral RNA, Molecular assay, specimens, S gene, SARS-CoV-2 RNA detection, Analysis, ORF1ab, target gene, acute respiratory syndrome, 95% CI, infected patients, clinical specificity, viral nucleic acid, Direct, objective, FIVE, Result, tested, performed, evaluate, significantly, assays, required, faster, were assessed, 【제목키워드】 COVID-19, Rapid, SARS-CoV-2 RNA,