Key Points:

  • PCRopsis™ Reagent RVD with RVD Enhancer is an alternative to RNA / DNA extraction for the amplification of nucleic acids
  • Offers comparable RT-qPCR / PCR results to extraction procedures for many applications 
  • Compatible with saliva, urine and nasopharyngeal / oropharyngeal swab specimens
  • Cost a fraction of traditional extraction procedures, with fewer steps
  • Compatible with most specimen transport mediums
  • Compatible with most leading RT-qPCR / PCR kits and master mixes
  • Compatible with automated sample processing equipment
  • Reagent RVD with RVD Enhancer is stored and shipped at room temperature​
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Intended Use (IN VITRO DIAGNOSTIC USE)

PCRopsis™ Reagent RVD with RVD Enhancer is intended for extraction-free amplification of RNA or DNA from properly collected and transported saliva or urine specimens or swab specimens in compatible transport mediums.

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  • Reagent RVD with RVD Enhancer may replace extraction procedures for many applications (e.g., life science research, veterinary pathogen research, etc.), but it must be verified by the end user.
Certificate of Analysis:
Lot017     Lot018     Lot020

PCRopsis™ Reagent RVD with RVD Enhancer is an alternative for:
  1. Qiagen: QIAamp Viral RNA mini Kit
  2. Qiagen: QIAmp DSP Viral RNA Mini Kit
  3. Qiagen EZ1 Advanced XL: EZ1 DSP Virus Kit
  4. Qiagen EZ1 Advanced XL: EZ1 Virus Mini Kit v2.0
  5. QIAGEN QIAcube: QIAmp DSP Viral RNA Mini Kit 
  6. QIAGEN QIAcube: QIAamp Viral RNA Mini Kit
  7. Roche MagNA Pure 24: MagNA Pure 24 Total NA Isolation Kit
  8. Roche MagNA Pure 96: DNA and Viral NA Small Volume Kit 
  9. Roche MagNA Pure LC: Total Nucleic Acid Kit 
  10. Roche MagNA Pure Compact: Nucleic Acid Isolation Kit I 
  11. bioMérieux NucliSENS® easyMAG®
  12. bioMérieux EMAG®
  13. Thermo Fisher / Applied Biosystems: MagMAX mirVana Total RNA Isolation Kit

​The Science Behind Reagent RVD with RVD Enhancer
PCRopsis™  Reagent RVD with RVD Enhancer is engineered to simultaneously bind a variety of RT-qPCR / PCR inhibitors found in clinical specimens, lyse microorganisms and stabilize RNA and DNA in a manner that's compatible with RT-qPCR / PCR. The product consist of a proprietary mixture of peptides, salts, stabilizers, buffers, sodium azide, and RVD Enhancer to achieve these tasks. Reagent RVD with RVD Enhancer allows for extraction-free amplification of microbial RNA / DNA without performing nucleic acid isolation, centrifugations or other sample manipulations which may introduce errors, contaminants and/or skew the representation of RNA fragments.
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User should verify the applicability of Reagent RVD with RVD Enhancer for unintended applications.

Reagent RVD with RVD Enhancer vs Old Fashion RNA Extraction

Traditional RNA Extraction Results in Significantly Different Levels of miRNA ... this bias can affect results.

Total RNA extraction from tissues for microRNA and target gene expression analysis: not all kits are created equal
Rikki A. M. Brown, Michael R. Epis, Jessica L. Horsham, Tasnuva D. Kabir, Kirsty L. Richardson & Peter J. Leedman BMC Biotechnology volume 18, Article number: 16 (2018)  - Link to manuscript

A comparison of RNA extraction and sequencing protocols for detection of small RNAs in plasma​
Ryan K.Y. Wong, Meabh MacMahon, Jayne V. Woodside & David A. Simpson BMC Genomics volume 20, Article number: 446 (2019)  - Link to manuscript

RNA Preservation Agents and Nucleic Acid Extraction Method Bias Perceived Bacterial Community Composition 
Ana McCarthy, Edna Chiang, Marian L. Schmidt, Vincent J. Denef, PLOS ONE (2015)  - Link to manuscript
Reagent RVD with RVD Enhancer does not use "capture & release" RNA / DNA technology (e.g., columns, beads, etc.), therefore it does not exhibit this bias.

Studies with PCRopsis™ Reagent RVD with RVD Enhancer
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Reagent RVD + RT-qPCR produces comparable results to RNA extraction (Qiagen) + RT-qPCR; even when samples are diluted 1/10 and 1/100 ​
RNA was extracted using Qiagen QIAamp Viral RNA Kit and samples processed with Reagent RVD according to manufacture's protocol. RT-qPCR was performed using IDT qPCR probe assay and Promega GoTaq® Probe 1-Step RT-qPCR System.
Reagent RVD + RT-qPCR amplifies a target gene fragment from 100% of tested samples containing a RNA virus (n=21) in UVT. This amplification results in comparable Ct values compared to RNA extraction from the same samples, even when the samples are diluted 1/10 or 1/100. Reagent RVD + RT-qPCR does not amplify the same target gene fragment in samples lacking the RNA virus (n = 34; data not shown).  35% of RNA virus positive samples not processed with Reagent RVD or RNA extracted, followed by RT-qPCR, were incorrectly identified as negative.
Reagent RVD + RT-qPCR detects 3.9 viral copies / µL from transport medium or saliva containing SARS-CoV-2​
Transport medium and human saliva were spiked with inactivated SARS-CoV-2 (ATCC) and processed with Reagent RVD with RVD Enhancer. Five viral gene sites were amplified through RT-qPCR, in duplicates, and averages shown. Reagent RVD with RVD Enhancer mediated amplification of tested gene regions with Ct values between 33 ~42 at 3.9 viral copies / µL​.
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RT-qPCR was performed using IDT qPCR probe assay and Promega GoTaq® Probe 1-Step RT-qPCR System.
Reagent RVD with RVD Enhancer results in lower Ct values with specimens in UVT, WHO VTM, CDC VTM or PBS as transport mediums
Various viral transport mediums (BD UVT, WHO VTM, CDC VTM, PBS) were spiked with SARS-CoV-2 and processed with Reagent RVD or Reagent RVD with RVD Enhancer. Reagent RVD with RVD Enhancer resulted in lower Ct values in all viral transport mediums. Reagent RVD alone produced Ct values comparable to extracted RNA when UVT was used as a transport medium. Reagent RVD with RVD Enhancer facilitates direct RT-qPCR from RNA viral samples in diverse transport mediums.

RNA was extracted using Qiagen QIAamp Viral RNA Kit and samples processed with Reagent RVD or Reagent RVD with RVD Enhancer according to manufacture's protocol. RT-qPCR was performed using IDT qPCR probe assay and Promega GoTaq® Probe 1-Step RT-qPCR System.
Reagent RVD + PCR has a limit of detection of <100 cfu / mL for gram positive and negative bacteria 
Reagent RVD facilitates extraction-free detection of gram positive and negative bacteria, with a limit of detection <100 cfu / mL in universal transport medium. 

Satisfaction Guarantee - Warranty Summary

Entopsis guarantees the performance of Reagent RVD with RVD Enhancer to facilitate the direct amplification of select viruses, bacteria and mammalian cells using PCR / RT-qPCR from saliva, urine and swab samples in compatible mediums when the sample is properly stored. Within 7 days of purchase, contact Entopsis (info@entopsis.com) should Reagent RVD with RVD Enhancer fail to perform as described above or in the CoA. In such a scenario, Entopsis will re-test the Lot in question using remaining reagent at our facility at our expense (purchaser does not need to ship product to Entopsis). If the re-tested Lot fails to meet our performance criteria, Entopsis will replace the faulty reagent free of charge. If the Lot meets our performance criteria as stated on the Certificate of Analysis for the Lot in question, then no additional action will be taken.

Please keep in mind that purchasers may use Reagent RVD with RVD Enhancer for numerous applications, sample types, gene targets, conditions, etc. that are untested and thereby may not work as desired. The purchaser must determine the suitability of Reagent RVD with RVD Enhancer for their particular use. However, the Entopsis scientific team can assist in the optimization of Reagent RVD with RVD Enhancer for your desired application.  Refunds will not be provided due to a lack of product compatibility with purchaser's desired application. This is why small test volumes of Reagent RVD with RVD Enhancer are offered. 

​THIS WARRANTY ONLY APPLIES IF PRODUCTS ARE PURCHASED FROM AN APPROVED VENDOR (E.G., ENTOPSIS).

Complete Terms and Conditions