Melting Curve-based Assay as an Alternative Technique for the Accurate Detection of SARS-CoV-2

• Published in: Advanced biomedical research Vol. 11; no. 1; p. 37
• Main Authors: Aboutalebian, Shima, Mirzaaghaei, Somaye, Fakhim, Hamed, Faramarzi, Sama, Mousavi, Somayeh, Ghafel, Safiyeh, Gholipour, Sahar, Farhang, Armin, Mirhendi, Hossein, Nikaeen, Mahnaz
• Format: Journal Article
• Language: English
• Published: India Wolters Kluwer - Medknow 01.01.2022; Wolters Kluwer Medknow Publications
• Subjects: diagnosis biological assay; e-gene; Original; rdrp-gene; sars-cov-2; sybr green
• ISSN: 2277-9175; 2277-9175
• DOI: 10.4103/abr.abr_87_21

Background: Early and cost-effective diagnosis and monitoring of the infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are critically important to anticipate and control the disease. We aimed to set up a SYBR Green-based one-step real-time polymerase chain reaction (PCR) as a lower-cost alternative method to detect the virus. Materials and Methods: An in-house SYBR Green-based PCR assay targeting the envelope (E) and RNA-dependent RNA polymerase (RdRp) genes, was set up to diagnose the infection, and was compared with the reference probe-based PCR method. Results: When the commercial probe-based assay was considered as the reference method, SYBR Green-based PCR had a slightly lower sensitivity (81.98% and 86.25% for E and RdRp targets, respectively) and a good specificity (100% and 94.44% for E and RdRp targets, respectively). For both gene targets, three different melting temperature (Tm) patterns were found in the PCRs of the nasopharyngeal/oropharyngeal swab samples, but no size polymorphism was seen in agarose gel electrophoresis. Conclusion: Further studies to improvement of the assay are needed to make it an inexpensive and reliable tool for the diagnosis of COVID-19.