QIAprep& Viral RNA UM Kit is an innovative liquid-based approach to viral RNA epidemiology. The kit is optimized for ultrafast sample preparation and sensitive real-time PCR detection of enveloped RNA viruses such as coronaviruses from human samples (nasal, oro-, and nasopharyngeal swabs) collected in non-fixation transport media like UTM, VTM, PBS, 0.9% NaCl, Virocult, and eSwab. Fewer and simpler workflow steps accelerate the turnaround time and increase testing frequency while decreasing plastic usage, cost, and hands-on time. The method can be seamlessly integrated into automated workflows with standard lab equipment at all throughput needs.
- Sample preparation and detection steps integrated into one single kit
- Time to result in under 1 hour
- End-to-end liquid-based workflow for full automation on liquid handlers
- Compatibility with any assay and real-time PCR cycler
- 4-plex (RT)-PCR capability for detection of up to 4 targets, including viral targets and/or controls
- Includes two controls (inhibition and sampling) for reliable interpretation of results
- Two-phase hot-start procedure and stable RT included in the master mix, for stability and specificity
The QIAprep& Viral RNA UM Kit can be used in conjunction with the SARS-CoV-2 N1+N2 Assay Kit.
The end-to-end liquid-based workflow of the QIAprep& Viral RNA UM Kit takes under one hour from start to finish. It encompasses an optional heat-treatment step and only three pipetting steps conducted directly in the PCR reaction vessel. This simplified procedure decreases the number of handling steps and plastic use and can be fully automated using liquid handlers. Simplicity and speed enable a throughput of up to 2600 samples per PCR cycler in an eight-hour shift (i.e., approximately seven runs of 384 samples each).
The kit is compatible with samples collected in non-fixation transport media like UTM, VTM, PBS, 0.9% NaCl, Virocult, and eSwab (see figure “Consistent results using six different transport media“).
Thanks to optimized chemistry, the new QIAprep& Viral RNA UM Kit delivers performance comparable to standard PCR workflows that use the gold standard for sample extraction (see figure “Reliable assessment of limit of detection (LOD) of viral particles“).
The QIAprep& Viral RNA UM Kit combines a liquid-based sample preparation step completed in only two minutes with real-time RT-PCR detection in a streamlined workflow. Users can automate this method with standard lab equipment for any throughput, assay and reaction need from single to multiplex testing.
The kit is compatible with dual-labeled probes, e.g., TaqMan® probes in multiplex one-step RT-PCR detection of one or more targets (altogether, up to 4 assays including the internal controls). The kit has been optimized for use with most real-time cyclers. A novel two-phase hot-start procedure ensures high specificity and sensitivity in real-time RT-PCR. For high in-process safety during virus detection, each kit contains reagents for the simultaneous detection of user-defined targets and two internal controls for confident result interpretation.
The first is the inhibition control that consists of a synthetic RNA with a unique and artificial sequence and its detection assay that can optionally be used to monitor successful amplification. The RNA IC is intended to report instrument or chemistry failures, errors in assay setup, and the presence of inhibitors. The second control is a sampling control, an assay that targets two human transcripts and can optionally be used to confirm that the primary sample tube contains human material and that RNA materials have not been degraded. Together, this allows correct interpretation of negative detection results.
|Assay||Target(s)||Reporter dye/color channel*||Supply|
|Inhibition Control||Synthetic IVT||Cy5/Red||Included in the kit, optional|
|Sampling Control||Human B2M and RNase P genes||HEX™/Yellow (two targets detected in the same channel)||Included in the kit, optional|
|Passive reference dye||–||ROX/Orange||Included in the kit, optional|
|Your viral RNA assay||User-defined||User-defined|
Recommended: target detection in green, blue, or NIR channel
|Provided by the user|
*Altogether, up to 4 assays or targets, including the internal controls and excluding the reference dye
The innovative QIAprep& offers a rapid and straightforward 3-step workflow: an aliquot is taken from a primary sample (nasopharyngeal, oropharyngeal, or nasal swab) in transport media as starting material. This can be subjected to an optional heat-treatment step before being added to an optimized sample preparation buffer directly in the PCR vessel, which allows the preparation of the viral RNA template without degradation in two minutes. Next, this is combined with an RT-qPCR reaction mix in the same tube or well for rapid amplification on any thermocycler using any assay. The output is finally interpreted, delivering test results in under one hour from start to finish – including incubation and hands-on time (see figure “An innovative 3-step liquid-based workflow”).
The QIAprep& Viral RNA UM Kit simplifies and accelerates PCR-based detection of enveloped RNA viruses such as SARS-CoV-2 coronavirus from human samples for epidemiological research purposes.