The Altona Diagnostics RealStar SARS-CoV-2 RT-PCR Kit 1.0 was the primary assay used for molecular diagnosis. A subset of SARS-CoV-2 positives were identified using the Cepheid Xpert Xpress SARS-CoV-2 GeneXpert platform per manufacturer instructions. All samples were processed within 24 hours of collection.
The RealStar SARS-CoV-2 RT-PCR Kit 1.0 total reaction volume was 30 μL (10 μL extracted sample and 20 μL Master Mix). The kit contains 2 premade master mixes, A and B, which contain PCR buffer, magnesium salt, primers and probes, reverse transcriptase, and DNA polymerase. The detectors used are Cy5 (SARS-CoV-2; S gene), FAM (B-βCoV; E gene), and JOE (Internal Control). CT values for the S gene target (Cy5) are reported inSupplemental Table 2 . Taqman RT-PCR was performed using the Prism 7500 Sequence Detection System (Applied Biosystems) at the following cycling conditions: 1 cycle at 55.0°C for 20 minutes, 1 cycle at 95.0°C for 2 minutes and 45 cycles at 95.0°C for 15 seconds, 55.0°C for 45 seconds and then 72.0°C for 15 seconds. Validation of this assay was performed as described in ref. 22 (link).
Testing on the Cepheid Xpert Xpress SARS-CoV-2 GeneXpert platform was performed in accordance with manufacturer’s instructions (40 ).
The RealStar SARS-CoV-2 RT-PCR Kit 1.0 total reaction volume was 30 μL (10 μL extracted sample and 20 μL Master Mix). The kit contains 2 premade master mixes, A and B, which contain PCR buffer, magnesium salt, primers and probes, reverse transcriptase, and DNA polymerase. The detectors used are Cy5 (SARS-CoV-2; S gene), FAM (B-βCoV; E gene), and JOE (Internal Control). CT values for the S gene target (Cy5) are reported in
Testing on the Cepheid Xpert Xpress SARS-CoV-2 GeneXpert platform was performed in accordance with manufacturer’s instructions (40 ).
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