The XpressAmp™ Direct Amplification Reagents provide a fast, RNA extraction-free method to prepare viral samples for PCR-based amplification using commonly available RT-qPCR reagents. Collect the samples by nasopharyngeal swab in universal or viral transport media, and perform direct amplification analysis in RT-qPCR. The simple sample preparation method requires only a 10-minute, room-temperature incubation that is easy to automate.

How the XpressAmp™ Direct Amplification Reagents Work

Direct PCR Amplification Benefits and Challenges

Viral RNA Detection Without the Need for Extraction

XpressAmp™ Detection of RNA from Inactivated Influenza/RSV Virus Pellet

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.

Amplification of Synthetic SARS-CoV-2 RNA from XpressAmp™ Lysates

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.

An independent laboratory performed an evaluation of the ability of the XpressAmp™ Lysis Buffer, used to prepare viral sample for RT-qPCR analysis, to inactivate the SARS-CoV-2 virus.

SARS-CoV-2 Viral Inactivation White Paper