NDLI: Detection of SARS-CoV-2 RNA by a Multiplex Reverse-Transcription Loop-Mediated Isothermal Amplification Coupled with Melting Curves Analysis

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Detection of SARS-CoV-2 RNA by a Multiplex Reverse-Transcription Loop-Mediated Isothermal Amplification Coupled with Melting Curves Analysis

Content Provider	MDPI
Author	Oscorbin, Igor Shevelev, Georgiy Pronyaeva, Ksenia Stepanov, Andrey Shamovskaya, Darya Mishukova, Olga Pyshnyi, Dmitrii Filipenko, Maksim
Copyright Year	2021
Description	Loop-mediated isothermal amplification (LAMP) is a method of nucleic acid amplification that is more stable and resistant to DNA amplification inhibitors than conventional PCR. LAMP multiplexing with reverse transcription allows for the single-tube amplification of several RNA fragments, including an internal control sample, which provides the option of controlling all analytical steps. We developed a method of SARS-CoV-2 viral RNA detection based on multiplex reverse-transcription LAMP, with single-tube qualitative analysis of SARS-CoV-2 RNA and MS2 phage used as a control RNA. The multiplexing is based on the differences in characteristic melting peaks generated during the amplification process. The developed technique detects at least 20 copies of SARS-CoV-2 RNA per reaction on a background of 12,000 MS2 RNA copies. The total time of analysis does not exceed 40 min. The method validation, performed on 125 clinical samples of patients’ nasal swabs, showed a 97.6% concordance rate with the results of real-time (RT)-PCR assays. The developed multiplexed LAMP can be employed as an alternative to PCR in diagnostic practice to save personnel and equipment time.
Starting Page	5743
e-ISSN	14220067
DOI	10.3390/ijms22115743
Journal	International Journal of Molecular Sciences
Issue Number	11
Volume Number	22
Language	English
Publisher	MDPI
Publisher Date	2021-05-27
Access Restriction	Open
Subject Keyword	International Journal of Molecular Sciences Analytical Chemistry Sars-cov-2 Coronavirus Loop-mediated Isothermal Amplification Lamp Multiplex Amplification Melting Curve Analysis
Content Type	Text
Resource Type	Article

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