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Detection of SARS-CoV-2 by Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Technique
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Ali Haji Esmaili Qomi1    , Mahtab Noorifard 2, Ramin Hamidi Farahani3 , Mehdi Shakori Khomartash3 , Abbas Morovvati4 , Ashkan Dirbazian4 , Pegah Shakib5 |
1- Infectious Diseases Research Center, Aja University Of Medical Sciences, Tehran, Iran.
2- Infectious Diseases Research Center, Aja University of Medical Sciences, Tehran, Iran , Mahtab-noorifard@yahoo.com
3- Infectious Diseases Research Center, Aja University of Medical Sciences, Tehran, Iran
4- Department of Microbiology, Faculty of Basic Science Qom Branch, Islamic Azad University, Qom, Iran
5- Razi Herbal Researches Center, Lorestan University of Medical Sciences, Khorramabad, Iran. |
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Abstract: (1004 Views) |
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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| Keywords: SARS-CoV-2, Covid-19, Reverse transcription loop-mediated isothermal a |
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Full-Text [PDF 383 kb]
(255 Downloads)
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Type of Study: Original Research |
Subject:
Microbiology
Received: 2021/09/3 | Accepted: 2022/05/10 | Published: 2023/09/27
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Ethics code: IR.AJAUMS.REC.1399.140
Haji Esmaili Qomi A, Noorifard M, Hamidi Farahani R, Shakori Khomartash M, Morovvati A, Dirbazian A et al . Detection of SARS-CoV-2 by Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Technique. SJKU 2023; 28 (4) :66-76 URL: http://sjku.muk.ac.ir/article-1-6937-en.html
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