The COVID-19 pandemic has highlighted the important role that polymerase chain reaction (PCR) can play in molecular diagnostics. PCR-based testing quickly became the predominant method used to identify infected individuals and stem chains of transmission. However, traditional PCR technologies depend on RNA or DNA extraction before PCR testing, which adds time and costs to the workflow.
Technology Networks spoke to Dr. George Huang, chief financial officer at Entopsis, to learn how these steps can be omitted using direct PCR technology. In this interview, Dr. Huang also discusses the advantages of this approach, why uptake has so far not been greater and what sets next generation direct PCR reagents apart.
Entopsis PCRopsis™ Reagent RVD-RT deliveries superior direct PCR performance compared to traditional direct PCR products at room temperature, according to a recent report.
Direct PCR, where the user does not perform RNA or DNA extraction before PCR, holds great promise for the PCR molecular diagnostics industry. Such approaches save time and money while streamlining protocols and potentially reducing errors. However, many direct PCR technologies fail to compare favorably with traditional RNA and DNA extraction procedures. This has impeded the widespread adoption of direct PCR technologies in many clinical laboratories.
PCRopsis™ direct PCR reagents solve this problem in a unique fashion. It uses proprietary nanotechnology to bind PCR inhibitors found in test samples, lyse viruses and cells, and stabilize RNA / DNA in a fashion that’s compatible with PCR. This novel approach offers superior performance compared to traditional direct PCR technologies.
Entopsis officially launches PCRopsis™ Reagent RVD-RT, that mediates room temperature direct PCR from saliva, urine, and swab samples in transport mediums. Furthermore, the company highlights key technological performance differences with a comparison to a market leader, Promega XpressAmp™ Direct Amplification Reagent.
PCRopsis™ Reagent RVD-RT offers a fast, low cost, and robust alternative to traditional RNA and DNA extraction. In contrast to other direct PCR solutions, Reagent RVD-RT produces PCR results that are comparable to traditional nucleic acid extraction approaches from swab and saliva samples. Moreover, Reagent RVD-RT is compatible with all tested transport mediums.
Full Press Release
PCRopsis™ Reagent SRVD was recently shown to mediate direct PCR amplification from viral, bacterial and mammalian targets from human saliva.
Reagent SRVD offers the PCR testing industry a robust, reliable, and cost-effective alternative to traditional RNA and DNA extraction for downstream PCR applications. You simply mix 1 volume of saliva with 1 volume of Reagent SRVD, heat at 95°C for 10 minutes, and then you use the processed sample directly into your PCR mixture.
“Our goal is to provide market leading direct PCR solutions to users of PCR technologies. The launch of Reagent SRVD expands our offering to saliva, a key diagnostic bio-fluid.”, said Obdulio Piloto, Ph.D., C.E.O. at Entopsis, Inc.
“We are listening to our customers. The release of Reagent SRVD for direct PCR applications from saliva is testament to that promise.” said Abhignyan Nagesetti, Ph.D., Head of Manufacturing at Entopsis, Inc.
The PCRopsis™ R&D team has been extremely diligent and successful in developing and validating direct PCR technologies for numerous applications. As such, we are expanding our business development team in the U.S., E.U. and Latin America.
Say tuned for future developments.
3/30/2021 0 Comments
PCRopsis™ Reagent RVD + PCRopsis™ RVD Enhancer offers a direct PCR solution for specimens in most non-inactivating transport mediums.
PCRopsis™ Reagent RVD alone has been repeatedly shown to facilitate extraction-free amplification of viral, bacterial, and mammalian specimens in universal viral transport (UVT) mediums, like BD™ Universal Viral Transport system. This is one of the most common specimen transport mediums in the U.S., although many other transport mediums are used in the U.S. and abroad.
Entopsis developed PCRopsis™ RVD Enhancer to expand the compatibility of PCRopsis™ Reagent RVD to process specimens in diverse transport mediums. PCRopsis™ Reagent RVD, when used in combination with PCRopsis™ RVD Enhancer, can process specimens in just about any non-inactivating transport medium. Compatible transport mediums now include viral transport mediums recommended by the Centers for Disease Control and World Health Organization, and even a simple phosphate buffered saline (PBS) solution.
“Reagent RVD + RVD Enhancer offers a competitive price and workflow advantage to any laboratory performing PCR-based testing”, said Pratik Sharma, VP of business development at Entopsis.
PCRopsis™ Reagent RVD was recently shown to mediate direct PCR amplification from viral and bacterial targets using low volumes of reagent and sample, thus drastically lowering costs and increasing testing accessibility.
Reagent RVD is a flexible direct PCR product for various research and clinical applications. The PCR testing industry is particularly price-conscious, with thin profit margins. As such, laboratories seek affordable and accurate solutions to offer clients the best services.
Recent miniaturization studies confirm Reagent RVD facilitates extraction-free RT-PCR and qPCR for less than $1.70 per reaction. Typically, RNA and DNA extraction kits cost $4.00 ~ $7.00 per extraction (2~4 times more expensive than Reagent RVD). Moreover, Reagent RVD even offers advantages over alternative direct PCR products currently on the market.
Reagent RVD advantages over alternative direct PCR products:
“It’s interesting how RNA and DNA extraction was mostly taken as a necessary first step in PCR applications for decades. It took the COVID-19 pandemic to get people questioning the validity of that presumption on a large scale. We see the development of affordable and robust direct PCR products as a means of making the PCR diagnostic industry more efficient and equitable”, said Obdulio Piloto, Ph.D., C.E.O. of Entopsis.
“Reagent RVD doesn’t just solve a key problem in the PCR industry, but also offers huge cost savings to partnering labs”, said Pratik Sharma, VP of business development at Entopsis.
2/22/2021 0 Comments
PCRopsis™ Reagent RVD-E allows for direct PCR amplification of test specimens from swabs without the need for transport mediums or lengthy RNA / DNA extraction protocols.
The first step in infectious disease testing involves swabbing a part of the body. This specimen-containing swab is placed in a tube with transport medium for shipping to a reference laboratory. The lab then performs ~10 steps to extract RNA or DNA from the transport medium for PCR testing.
The problem with this approach is that it requires many consumables that present testing bottlenecks at various points; if any are not available, then testing cannot be performed. Moreover, the multi-step process is laborious and unnecessarily error-prone; every consumable and step presents an opportunity for misdiagnosis.
What if we could skip the need for transport medium and RNA / DNA extraction?
PCRopsis™ Reagent RVD-E simplifies the testing process while saving time and lowering costs. In this new scenario, specimen-containing swabs are shipped to a testing lab in an empty transport tube. The lab then simply adds a small amount of PCRopsis™ Reagent RVD-E, vortexes the mixture and heats it. The heated sample is directly applied into downstream PCR applications.
“PCRopsis™ Reagent RVD-E will impact testing paradigms beyond COVID-19, and increase the efficiency of various other tests, including respiratory panels, wounds and drug resistance”, said Francis Lim, Ph.D., senior scientist at Entopsis.
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Reagent RVD has proven capable of amplifying SARS-CoV-2 gene targets directly from universal viral transport (UVT) medium without the need for viral RNA extraction. However, diagnostic reference laboratories also want to use this approach for the amplification of bacterial gene targets.
The PCRopsis™ research team has extended their validation efforts in this direction. It’s been confirmed that Reagent RVD is capable of mediating extraction-free qPCR of specimens containing gram positive and negative bacteria.
To date, Reagent RVD facilitates extraction-free amplification of viral, bacterial and human gene targets directly from UVT.
This opens the door to new applications with Reagent