Qiagen, 221517, QIAprep& Buffer AB (16.4 ml)

For the preparation of 2400 samples requiring pre-treatment, including but not limited to saliva, gargle and sputum

Features

- Sample preparation and real-time PCR-based virus detection integrated into one single kit
- Time to result in under 1 hour
- End-to-end liquid-based workflow for complete 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

Product Details

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. The kit is also compatible with neat saliva and gargle samples. 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 QIAsymphony Kit can be used for automated sample preparation and assay set up on the QIAsymphony SP/AS. The QIAprep&amp Viral RNA QIAsymphony Kit requires the installation of a custom protocol available from QIAGEN Services.

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

Want to try the QIAprep&amp Viral RNA UM Kit for the first time? Register to start your trial .

For labs that need to track currently relevant mutations of the virus, wet-lab tested LNA-based qPCR genotyping assays have been designed to detect the variants of concern (VOCs). The assay performance in terms of specificity and sensitivity has been verified using the QIAprep&amp Viral RNA UM Kit. View the assays here .

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 real-time 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 "). The kit is also compatible with neat saliva and gargle samples (See figures " QIAprep&amp Viral RNA UM Kit performance comparison between saliva and gargle samples " and " Protocol comparison of QIAprep&amp Viral RNA UM Kit performance with saliva samples ").

Thanks to optimized chemistry, the new QIAprep&amp Viral RNA UM Kit delivers performance comparable to standard qPCR workflows that use the gold standard for sample extraction (see figure " Reliable assessment of limit of detection (LOD) of viral particles ").

Principle

Assay: Target(s)
Inhibition Control: Synthetic IVT
Sampling Control: Human B2M and RNase P genes
Passive reference dye: -
Your viral RNA assay: User-defined

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.

*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 for virus detection: 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 two minutes. For saliva and gargle samples, we have developed a dedicated protocols that includes a short heat pretreatment step. 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 real-time PCR-based detection of enveloped RNA viruses such as SARS-CoV-2 coronavirus from human samples for epidemiological research purposes.