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

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Article Type:
Research/Original Article (دارای رتبه معتبر)
Abstract:
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.

Language:
Persian
Published:
Scientific Journal of Kurdistan University of Medical Sciences, Volume:28 Issue: 4, 2023
Pages:
66 to 76
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