(1056-A) Automated workflow from sample lysis to amplification for rapid SARS-CoV-2 detection

Abstract: When detecting nucleic acids, two main steps must be carried out, both of which can be very laborious and time-consuming: Firstly, the multi-step extraction of the nucleic acids from the sample material and secondly, the amplification and detection of the extracted nucleic acids, which is usually done by real-time PCR.
With the LumiraDx SARS-CoV-2 RNA STAR Complete Kit, the nucleic acid extraction is reduced to a single step where the samples or controls are mixed with an extraction buffer. Furthermore, by using the innovative and patented nucleic acid amplification technique qSTAR (qualitative selective temperature amplification reaction), the time to amplification is reduced to only 20 minutes by drastically reducing the temperature delta (~57°C - ~63°C) during the reaction. In this process, an additional nicking enzyme cleaves the newly formed DNA within the temperature cycles, generating more target DNA template while the polymerase works in parallel. The CyBio FeliX automates the LumiraDx assay in sub-steps that can be performed individually, as a complete run or modularly via the user-friendly Application Studio software. After transfer of the prepared and sealed reaction plates into the qTOWER3 real-time thermal cycler, the target sequence of SARS-CoV-2 (OFR1a) and the internal control sequence are detected in the FAM and ROX channels, respectively. In a direct comparison, automatically processed samples were compared with manually processed samples on one plate.
The automated preparation of the plate with extraction buffer and reaction mixture by the Analytik Jena CyBio FeliX led to homogeneous results in the same range as those of the manually prepared wells. The target sequence was only detected in wells with positive control medium, while the control sequence was detected in all wells.
Overall, the LumiraDx SARS-CoV-2 RNA STAR complete kit minimises the time-consuming extraction from swab samples to only one step and enables very rapid amplification of target sequences. Process automation on the Analytik Jena CyBio FeliX also enables flexible and customised maximisation of throughput. Through the Application Studio software, each automated process step can be executed separately. Amplification with the qTOWER3 is also efficient, highly reproducible and delivers homogeneous results across the entire reaction plate.