Key Points:

• PCRopsis™ Reagent SRVD 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 100% saliva
• Cost a fraction of traditional extraction procedures, with fewer steps
• Compatible with most leading RT-qPCR / PCR kits and master mixes
• Compatible with automated sample processing equipment
• Reagent SRVD is stored and shipped at room temperature

​Intended Use (IN VITRO DIAGNOSTIC USE)

• PCRopsis™ Reagent SRVD is intended for extraction-free amplification of RNA or DNA from properly collected and transported saliva samples.
• Reagent SRVD 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.

​The Science Behind Reagent SRVD

PCRopsis™ Reagent SRVD is engineered to simultaneously bind a variety of RT-qPCR / PCR inhibitors found in saliva 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 and buffers to achieve this task. Reagent SRVD 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 SRVD for unintended applications.

​Product Deterioration

PCRopsis™ ​Reagent SRVD should not be used if (1) there is evidence of damage or contamination to the product, (2) there is evidence of leakage, (3) the color of the reagent has changed from clear-white hazy, (4) the expiration date has passed, or (5) there are other signs of deterioration.​

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 SRVD does not use "capture & release" RNA / DNA technology (e.g., columns, beads, etc.), therefore it does not exhibit this bias.

​Extraction-Free Protocol

Test Sample:

•  100% saliva
• the compatibility of saliva in specific transport mediums needs to be confirmed 

Optional: replace 'Reagent SRVD' in the protocol below with a mixture of 5 parts Reagent SRVD + 2 parts PCRopsis™ RVD Enhancer

• ​This mixture may result in lower Ct values, ability to use various transport mediums and lower costs

1. Thoroughly mix Reagent SRVD to ensure homogeneity, but avoid creating bubbles unnecessarily 
   1. Reagent SRVD has a hazy, white color when homogenized and normal settlement occurs if not regularly mixed
2. Mix 1 volume of sample (20 μL) with 1 volume of Reagent SRVD (20 μL) in a thin walled tube (0.2 ~ 0.6 mL) or 96-well PCR plate
3. Pipette up & down to ensure complete mixing and then cap tube or apply plate sealer to plate to prevent evaporation
4. Heat diluted sample at 95°C and let cool at room temperature for ~10 seconds before continuing  
   1. Recommended heating times at 95°C:
      1. Mammalian: 5 minutes
      2. Viruses:         10 minutes
      3. Bacteria:       15 minutes
      4. NOTE: heating for a longer period of time does not negatively affect results
   2. Make sure the heating device has reached the desired temperature before applying sample
   3. Sample heating can be performed using a controlled heating block or thermal cycler
5. Mix heated sample and use 5 - 10 μL of lysed / stabilized sample in your desired RT-qPCR procedure
   1. ​Lysed sample can represent 25%~50% of your final RT-qPCR mixture (i.e., 5 ~ 10 μL sample into a total volume of 20 μL)

Limitations of the Procedure

• Performance characteristics of Reagent SRVD were validated using SARS-CoV-2, S. aureus and P. aeurginos spiked into human saliva. The use of alternative microorganisms, gene targets and / or detection methods may affect the performance of the product.
•  Repeated freezing and thawing of test specimens may reduce the detection of desired gene targets.
• Follow recommended guidelines for specimen collection as this may affect the ability to detect desired organisms.

PCRopsis Reagent SRVD is an alternative for:

1. Thermo Fisher Scientific: various MagMAX kits
2. ​Qiagen: RNeasy Saliva Kit, DNeasy Blood & Tissue Kit, etc.
3. Numerous other silica- or bead-based RNA and DNA extraction kits

​Performance & CoA

PCRopsis™ Reagent SRVD enables consistent amplification of RNA and DNA gene targets derived from viruses (e.g., coronaviruses), bacteria (e.g., S. aureus), and mammals (e.g., human) without the need for nucleic acid extraction procedures. Each lots is carefully tested for sterility and amplification of RNA or DNA to ensure quality and consistent results.

Certificate of Analysis:
LOT4    

Storage

This product is ready for use and no further preparation is necessary. The product should be transported and stored in its original container at 4–25°C until used. Do not overheat. Do not incubate or freeze prior to use. Improper storage will result in a loss of efficacy. Do not use after expiration date, which is clearly indicated on the bottle.
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Warnings & Precautions

FOR IN VITRO DIAGNOSTIC USE 

• Observe approved biohazard precautions and aseptic techniques to prevent contamination of the product. To be used only by adequately trained and qualified personnel.
• Pathogenic microorganisms, including hepatitis viruses and Human Immunodeficiency Virus, may be present in clinical specimens. "Standard Precautions" and institutional guidelines should be followed in handling all potentially bio-hazardous materials.
• Sterilize all biohazard waste including specimens, containers and mediums after their use.
• Directions should be read and followed carefully.
• Do not re-pack.
• The use of this product in association with a rapid diagnostic kit, diagnostic instrumentation or used in a manner not intended should be validated by the user.
• Do not ingest the reagent.
• Avoid skin contact with reagent since it contains sodium azide to prevent microbial growth.

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Studies with PCRopsis™ Reagent SRVD

Stay Tuned

Satisfaction Guarantee - Warranty Summary

Entopsis guarantees the performance of Reagent SRVD to facilitate the direct amplification of coronaviruses using RT-qPCR from saliva and the sample is properly stored. Within 7 days of purchase, contact Entopsis (info@entopsis.com) should Reagent SRVD fail to perform as described above or in the Certificate of Analysis (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 CoA for the Lot in question, then no additional action will be taken.

Please keep in mind that purchasers may use Reagent SRVD 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 the Reagent SRVD for its particular use. However, the Entopsis scientific team can assist in the optimization of Reagent RVD 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 SRVD are offered. 

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

Complete Terms and Conditions

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