Detection of SARS-CoV-2 by Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Technique

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Background and Aim: Covid-19 disease is a global health concern that requires rapid quantitative diagnostic testing.The standard technique for diagnosing SARS-CoV-2 is now REAL-TIME PCR.The aim of this study was to design a sensitive Reverse Transcription Loop-Mediated Isothermal Amplification method for the early identification of SARS-CoV-2. Material and Methods:In this laboratory study, specimens were prepared from the patients by laryngeal, pharyngeal, and nasal swabs and after RNA extraction and cDNA synthesis.SARS-CoV-2 virus was evaluated using the RT-LAMP method.Sensitivity of the RT-LAMP method was evaluated by primers targeting N and E genes.Results: In this study, 19 SARS-CoV-2 RNA samples were examined.According to the results of this study turbidity or discoloration of fluorescent dye showed that agarose electrophoresis gel is a more sensitive test.Detection limits for the number of E gene copy were observed at the dilutions of 10 -9 and 10 -8 for gel electrophoresis and fluorescent light, respectively.This means detection of 1 copy and 14 copies for the two above-mentioned methods, respectively.However, in the case of the N gene the detection limits, were at the dilutions of 10 -10 on the gel and 10 -8 by fluorescent light, which indicated detection of 1 copy and 17 copies, respectively.The calculations were performed using the Chiang formula. Conclusion:The results of this study showed that the LAMP was a simple, fast, sensitive, and specific method for the diagnosis of SARS-CoV-2 that may improve the diagnostic potential in clinical laboratories.

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