Evaluation and optimization of the SARS-CoV۲ genomic RNA extraction

BACKGROUND AND OBJECTIVESThe new coronavirus (SARS-CoV۲) is the cause of Covid-۱۹, and according to the WorldHealth Organization (WHO), has been a global problem and until last year, as a newepidemic, had covered all the countries of the world. This virus has a positive RNA of ۲۹.۸kb. The most important diagnostic method for the new coronavirus is based on genome-baseddetection or real-time RT-PCR. For this purpose, specific parts of RNA virus are identified.Therefore, the first step for identification is the viral genome (RNA) extraction. In theseviruses, extracting the genome with appropriate quality will be necessary for success inmolecular RT-PCR methods. Therefore, various and sometimes field and mobile methods areused for the genome extraction of pathogens . The aim of this work is a simple and portableRNA extraction method for detection of SARS-CoV۲ in the field.MATERIALS AND METHODSPharyngeal contaminated samples with SARS-CoV-۲ were obtained from Chamran Hospitaland were initially confirmed using a specific test (real-time PCR). Then the genome (RNA)was extracted using filtered syringe and ImmunoMagnetic Separation (IMS) methods from acertain concentration of the infected sample containing SARS-CoV-۲. Finally, theperformance and sensitivity of the mentioned method in genome extraction were investigatedusing agarose electrophoresis, real-time RT-PCR, and RT-LAMP.RESULTS AND DISCUSSIONThe data indicated that the efficiency of two manual methods (filtered syringe and IMS) inextracting SARS-CoV۲ viral genome can compete with commercial kits based on columnsand centrifuges.CONCLUSIONIn conclusion, the innovative methods proposed in this research can be used for field andmobile extraction of viral genomes in order to perform molecular detection tests

کلیدواژه ها:

Genome extraction ، New coronavirus ، Filtered Syringe ، Magnetic Nanoparticles ، Detection.