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QIAprep&amp Viral RNA UM Kit

For rapid and high-frequency PCR-based SARS-CoV-2 detection for epidemiological research
  • 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&amp 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.

 

The QIAprep&amp Viral RNA UM Kit can be used in conjunction with the SARS-CoV-2 N1+N2 Assay Kit.

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Cat No./ID: 221415
QIAprep&amp Viral RNA UM Kit (600)

For 600 x 20 µl reactions: 1.2 ml Viral RNA UM Prep Buffer, 2x 1.5 ml Viral RNA Master Mix, 4x, 1.2 ml RNA IC Template + Assay, 0.6 ml Human Sampling IC Assay, 1ml QN ROX, 2x 1.9 ml RNase-Free Water

Cat No./ID: 221417
QIAprep&amp Viral RNA UM Kit (2400)

For 2400 x 20 µl reactions: 4x1.2 ml Viral RNA UM Prep Buffer, 8x 1.5 ml Viral RNA Master Mix, 4x, 4x1.2 ml RNA IC Template + Assay, 4x0.6 ml Human Sampling IC Assay, 4x1 ml QN ROX, 8x 1.9 ml RNase-Free Water

The QIAprep&amp Viral RNA UM Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.


Product Details

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An innovative 3-step liquid-based workflow
An aliquot is taken from a primary sample (nasopharyngeal, oropharyngeal or nasal swab) in transport media as starting material. This is added to an optimized sample preparation buffer, which allows preparation of the viral RNA template without degradation in under two minutes, before being combined with an RT-qPCR reaction mix for rapid amplification on any thermocycler using any assay. The output is finally interpreted, delivering test results in under one hour.
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Reliable assessment of limit of detection (LOD) of viral particles
Limit of Detection (LOD) tested on inactivated SARS-CoV-2 viral particles at 250, 500, 1000, 2000 and 4000 cp/ml. Type of sample diluted in UTM + cell line KG1A at different cp/reaction. All samples tested with KG1A (cell line) amount into the reaction: 1200 cells per reaction. LOD of 8 copies/reaction offers the test sensitivity required.
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Six transport media compatible with the kit
The kit is compatible with samples collected in non-fixation transport media like UTM, VTM, PBS, 0.9% NaCl, Virocult and eSwab. 8 µl of respective transport medium spiked with indicated amount of IVT + 2 µl Viral RNA UM Prep Buffer + 10 µl Viral RNA UM Master Mix show comparable Ct values.
Performance

The end-to-end liquid-based workflow of the QIAprep&amp 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&amp 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").

Principle

The QIAprep&amp 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

Procedure

The innovative QIAprep&amp 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 preparation of the viral RNA template without degradation in under 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”).

Applications

The QIAprep&amp 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.

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