Abstract
Nucleic acid tests to detect the SARS-CoV-2 virus have been performed worldwide since the beginning of the COVID-19 pandemic. For the quality assessment of testing laboratories and the performance evaluation of molecular diagnosis products, reference materials (RMs) are required. In this work, we report the production of a lentiviral SARS-CoV-2 RM containing approximately 12 kilobases of its genome including common diagnostics targets such as RdRp, N, E, and S genes. The RM was measured with multiple assays using two different digital PCR platforms. To measure the homogeneity and stability of the lentiviral SARS-CoV-2 RM, reverse transcription droplet digital PCR (RT-ddPCR) was used with in-house duplex assays. The copy number concentration of each target gene in the extracted RNA solution was then converted to that of the RM solution. Their copy number values are measured to be from 1.5 × 105 to 2.0 × 105 copies/mL. The RM has a between-bottle homogeneity of 4.80–8.23% and is stable at 4 °C for 1 week and at −70 °C for 6 months. The lentiviral SARS-CoV-2 RM closely mimics real samples that undergo identical pre-analytical processes for SARS-CoV-2 molecular testing. By offering accurate reference values for the absolute copy number of viral target genes, the developed RM can be used to improve the reliability of SARS-CoV-2 molecular testing.
Highlights
The severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) that originated in Wuhan, China, in 2019 causes COVID19 infections [1] and can lead to severe lung damage and mortality [2]
While antigen and antibody testing kits have been developed for the rapid diagnosis of COVID-19, the World Health Organization (WHO) recommends the use of nucleic acid tests (NATs) as a standard means of confirmation of SARS-CoV-2 infection due to
Various reference materials (RMs) for SARS-CoV-2 are currently available; most are limited as users prefer RMs with reference values in absolute quantification of specific genes rather than total nucleic acid measurements or relative quantifications
Summary
The severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) that originated in Wuhan, China, in 2019 causes COVID19 infections [1] and can lead to severe lung damage and mortality [2]. While antigen and antibody testing kits have been developed for the rapid diagnosis of COVID-19, the World Health Organization (WHO) recommends the use of nucleic acid tests (NATs) as a standard means of confirmation of SARS-CoV-2 infection due to. The importance of reference materials (RMs), meaning homogenous and stable materials with specified properties [6], has been highlighted as molecular diagnostics are widely used in diagnosing infectious diseases worldwide [7]. RMs for specific pathogens are needed for the accurate quantification of their genetic materials by setting reliable values to compare and evaluate the performance of diagnostic kits and to compensate human errors during the process. Various RMs for SARS-CoV-2 are currently available; most are limited as users prefer RMs with reference values in absolute quantification of specific genes rather than total nucleic acid measurements or relative quantifications. The positive and negative RMs were dispensed in 1-mL aliquots into 1.8-mL Cryotubes (Thermo Fisher, Waltham, MA) and stored at − 70 °C
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
