Abstract
The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomeme SARS-CoV-2 test, and the Precision Biomonitoring TripleLock SARS-CoV-2 test on a field-deployable point-of-care real-time PCR instrument, Franklin three9, in combination with Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0 (M1) manual extraction system for rapid, specific, and sensitive detection of SARS-COV-2 in cell culture, human, and animal clinical samples. The Biomeme SARS-CoV-2 assay, which simultaneously detects two viral targets, the orf1ab and S genes, and the Precision Biomonitoring TripleLock SARS-CoV-2 assay that targets the 5′ untranslated region (5′ UTR) and the envelope (E) gene of SARS-CoV-2 were highly sensitive and detected as low as 15 SARS-CoV-2 genome copies per reaction. In addition, the two assays were specific and showed no cross-reactivity with Middle Eastern respiratory syndrome coronavirus (MERS-CoV), infectious bronchitis virus (IBV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, and other common human respiratory viruses and bacterial pathogens. Also, both assays were highly reproducible across different operators and instruments. When used to test animal samples, both assays equally detected SARS-CoV-2 genetic materials in the swabs from SARS-CoV-2-infected hamsters. The M1 lysis buffer completely inactivated SARS-CoV-2 within 10 min at room temperature enabling safe handling of clinical samples. Collectively, these results show that the Biomeme and Precision Biomonitoring TripleLock SARS-CoV-2 mobile testing platforms could reliably and promptly detect SARS-CoV-2 in both human and animal clinical samples in approximately an hour and can be used in remote areas or health care settings not traditionally serviced by a microbiology laboratory.
Highlights
The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission
The Biomeme SARS-CoV-2 test allows for the specific detection of SARS-CoV-2 Open Reading Frame 1ab (Orf1ab) and Spike (S) genes, while an RNA Process Control spiked into the sample (RNA extraction and RT-PCR control utilizing MS2 bacteriophage), similar to RNaseP below, must be detected for negative SARS-CoV-2 result to be valid
To safely use the Biomeme and Precision Biomonitoring SARS-CoV-2 Test kits at the point-of-care, it is critical to demonstrate a complete inactivation of SARS-CoV-2 by the lysis buffer in the M1 extraction cartridge
Summary
The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. The 5′ UTR contains stem-loop structures that play a functional role in viral replication, whereas the E gene is involved in virus assembly, budding, and pathogenesis[5,6] This test uses human RNaseP, a ubiquitously expressed gene in all human cells[7], as the third target to serve as an internal control for RNA extraction and PCR amplification. This study describes both Biomeme SARS-CoV-2 and the Precision Biomonitoring TripleLock SARS-CoV-2 tests on Franklin three[9], a portable real-time PCR instrument that has been used to successfully detect foot‐and‐mouth disease[8] and African swine fever[9] viruses in clinical samples. We describe the performance characteristics of each assay using clinical and animal samples and demonstrate the potential applications for settings outside of a traditional microbiology laboratory
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