Simplified Magnetic Bead Based Viral RNA Extraction for Rapid Detection of COVID-19

Shashi Sharma,Deepak Pardasani, Pooja Yadav, Jyoti S Kumar,Suman Dhankher, Ambuj Srivast, Sushil K Sharma,Devendra Kumar Dubey,Paban Kumar Dash

Research Square (Research Square)(2021)

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摘要
Abstract Rapid and large-scale diagnosis has helped in mitigation the recent ongoing pandemic of corona virus disease of 2019 (COVID-19). The pandemic had a devastating effect on global economy. The molecular detection system has evolved over last two decades and is rapidly replacing the conventional confirmatory techniques in diagnostic virology. However the major limitation in implementation of available molecular detection assays is the non availability of field deployable nucleic acid isolation platform. The standard laboratory diagnosis rely on confirmation of presence of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in respiratory specimens of suspected patients. Preparation of viral nucleic acid is a critical step involved followed by downstream molecular diagnostic platforms. For good quality of viral RNA extraction many commercial extraction kits, are available. These are developed in a surge of pandemic scenario keeping in view the large demand for testing. The commercial RNA extraction kits available on either column based or magnetic extraction are limited and, alternative, non-commercial protocols are rapidly required. Here, we have standardized an in-house magnetic bead RNA extraction method which utilises simple in-house reagents and manual extraction method that doesn’t require any high-end equipments. The in-house assay was evaluated against the commercial available silica column and magnetic extraction kits using a panel of 100 throat /nasal swab samples. A high correlation in viral RNA detection with TaqMan qRT-PCR was observed with excellent sensitivity and specificity. Interestingly, the developed method is very simple, cost effective, rapid and can be quickly add up any downstream amplification platform for SARS-CoV-2 detection.
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rna,extraction,rapid detection
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