Direct PCR: XpressAmp® Direct Amplification Reagents

Viral transport media (VTM) was inoculated with a nasopharyngeal swab and spiked with RSV A and Influenza B (Hong Kong) virus reconstituted from Helix Elite™ Inactivated Standard, Inactivated Influenza A/B and Respiratory Syncytial Virus. This high-level virus sample (1 × 10³ copies/μl) was diluted 1:10 and 1:100 in VTM to create the medium- and low-level virus samples. In parallel, two users created sample lysates from the spiked VTM samples using the XpressAmp® Direct Amplification Reagents. Both users then detected the presence of RSV A and Influenza B by RT-qPCR using GoTaq® Probe 1-Step qPCR System (Cat.# A6121) supplemented with the XpressAmp® Solution. N=6.

Viral transport media (VTM) was inoculated with a nasopharyngeal swab and spiked with Synthetic SARS-CoV-2 RNA Control 2 (Twist Biosciences, Cat.# 102024, final concentration 1 × 10⁴ copies/μl). Spiked VTM samples were lysed by combining 5μl of sample with 5μl of prepared XpressAmp® Lysis Buffer and incubated at room temperature for 10 minutes. Following incubation, 5μl of sample lysate was added to a monoplex GoTaq® Probe 1-Step RT-qPCR (25μl) containing XpressAmp® Solution and amplified using the 2019 nCoV RUO kit (IDT, Cat.# 10006713) and thermal cycled according to the CDC protocol. N=8 amplification replicates.