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VLP-RNA Extraction Control Orange

Molecular IVD companies continue to experience tighter and tighter regulations in regard to incorporating appropriate positive and negative controls into their assays. Meridian’s VLP-RNA Extraction Controls offer a simple and effective way to reduce false-negative results in RT-qPCR assays. The biomimetic controls are composed of virus-like particle (VLP) shells packaged with a defined number of copies of target RNA molecules. Their composition closely resembles a test sample and serves as a full-process quantitative PCR positive control, monitoring the success of an RT-qPCR assay from lysis/extraction to reverse transcription and amplification.

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Meridian's VLP-RNA Extraction Control Orange

  • Contains a defined number of copies of target RNA molecules, encapsidated within a virus-like particle (VLP).

  • Closely mimics the test sample, undergoing the same processing from lysis and extraction to RT-qPCR detection.

  • Non-infectious material for ease of handling and shipping.

  • Compatible with commonly used RNA extraction methods and lyophilization for creating freeze-dried mixes.

Why use VLP-RNA Extraction Controls in an RT-qPCR assay?

Possible causes of negative detection in an RT-qPCR
VLP-RNA Extraction Controls graph
Spike-In internal controls are RNA molecules added between the nucleic acid isolation and the detection, they do not monitor the extraction and lysis steps. Mimics internal controls are RNA molecules provided in a format resembling a biological sample (i.e. micelles) and are added just before lysis, but do not allow long incubation with the sample. VLP-RNA Extraction Control have better nuclease resistance, for long incubation with the sample, better quantification for consistent input in MDx processes and are easier to ship and store, ensuring that the MDx process provides reliable results by questioning each step of the process from lysis, through extraction to detection.

VLP-RNA Extraction Control Orange, MDX069

Available in 1 mL (~1×104 copies/µL) and 20 mL (~1×104 copies/µL) aliquots

Description

VLP-RNA Extraction Control Orange contains an internal control RNA sequence, with no known homology to any organism, encapsidated in a virus-like particle. The VLP-RNA Extraction Control is spiked in the sample prior to RNA extraction. Following RNA extraction, VLP-RNA Extraction Control can be detected in RT-qPCR by adding the VLP Detection Mix (containing specific primers and probe (emission wavelength = 560nm)) to the reaction mix. The detection of VLP-RNA Extraction Control acts as a PCR positive control and confirms the success of the extraction, reverse transcription, and amplification steps, avoiding the misinterpretation of a false negative result.


Specifications

Description An Internal Control RNA sequence, with no known homology to any organism, encapsidated in a virus-like particle and a VLP Detection Mix containing primers and probe to the internal control RNA sequence, used to confirms the success of the extraction, reverse transcription, and amplification steps
Concentration 104 copies/µL
Detection Mix Primers and HEX labeled probe (absorption λmax = 535 nm and emission λmax = 555 nm)
Application one-step RT-qPCR
Sample type Tissue, cells
Presentation 2 vials
Storage -20 °C
Mix stability See outer label
Functionality test VLP Detection Mix tested in an RT-qPCR of a serial dilution of Internal Control RNA.

FAQs: VLP-RNA Extraction Control

When performing RNA extraction, it is often advantageous to have an exogenous source of RNA template that is spiked into the sample prior to lysis. This control RNA is then co-purified with the sample RNA and can be detected as a PCR positive control for the whole extraction process.

Although it is not mandatory yet, regulators across the world are tightening up recommendations for the addition of internal controls, EU: IVDD to IVDR 2017/746 “Control materials with quantitative or qualitative assigned values intended for one specific analyte or multiple analytes shall be classified in the same class as the device.” MDSAP with FDA (“general controls ensuring effectiveness of the device”), TGA (“provisions for the user, at time of use, of device performance”), MHLW and ANVISA

The VLP-RNA Extraction Controls are in plant based VLPs, so there is no animal/human viruses used within the process, so there is no possibility of the VLP-RNA Extraction Control giving false positive results.

The VLP-RNA Extraction Controls are extremely robust and stable quantitative PCR positive control, we have tested then for 20 cycles of freeze thawing, storage at ambient temperature for one month, heating to 50 ᵒC for 10 minutes, lyophilization and nuclease treatment without seeing any change in Ct values.

The VLP-RNA Extraction Controls are in plant based VLPs, so there is no animal/human viruses used within the process, so there is no possibility of the VLP-RNA Extraction Control giving false positive results.

The primers and probe for the VLP-RNA Extraction Control are in the VLP Detection Mix for VLP-RNA Extraction Control Red, MDX068 and VLP-RNA Extraction Control Orange, MDX069. However we do not supply the primers and probe for the custom VLP-RNA Extraction Control, MDX071.

The two colors are for the fluorescent dyes on the VLP-RNA Extraction Control probe. VLP Detection Mix Red (Cy5 – emission wavelength = 670nm) and VLP Detection Mix Orange (HEX – emission wavelength = 555nm). This allows you to chose one that will fit with your existing protocol in a multiplex RT-qPCR assay.

No, the VLP-RNA Extraction Control will work with commercially available silica-membrane extraction kits and CHELEX matrices and has been tested on a wide range of qPCR platforms.

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Have questions about a product? Want to learn more about Meridian’s molecular or immunoassay reagent portfolio? We want to hear from you!


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