Author/Authors :
Aboutalebian, Shima Department of Medical Parasitology and Mycology - School of Medicine - Research Core Facilities Laboratory - Isfahan University of Medical Sciences, Isfahan, Iran , Mirzaaghaei, Somaye Department of Medical Parasitology and Mycology - School of Medicine - Research Core Facilities Laboratory - Isfahan University of Medical Sciences, Isfahan, Iran , Fakhim, Hamed Infectious Diseases and Tropical Medicine Research Center - Isfahan University of Medical Sciences, Isfahan, Iran , Faramarzi, Sama Department of Medical Parasitology and Mycology - School of Medicine - Research Core Facilities Laboratory - Isfahan University of Medical Sciences, Isfahan, Iran , Mousavi, Somayeh Department of Medical Parasitology and Mycology - School of Medicine - Research Core Facilities Laboratory - Isfahan University of Medical Sciences, Isfahan, Iran , Ghafel, Safiyeh Department of Medical Microbiology - School of Medicine - Isfahan University of Medical Sciences, Isfahan, Iran , Gholipour, Sahar Department of Environmental Health Engineering - Faculty of Health - Kashan University of Medical Sciences, kashan, iran , Farhang, Armin Department of Pharmaceutical Biotechnology - School of Pharmacy - Isfahan University of Medical Sciences, Isfahan, Iran , Mirhendi, Hossein Department of Medical Parasitology and Mycology - School of Medicine - Research Core Facilities Laboratory - Isfahan University of Medical Sciences, Isfahan, Iran , Nikaeen, Mahnaz Department of Environmental Health Engineering - School of Health - Isfahan University of Medical Sciences, Isfahan, Iran
Abstract :
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.
Keywords :
E‐gene , RdRp‐gene , SARS‐CoV‐2 , SYBR Green , Diagnosis Biological Assay
