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Powerful, versatile SARS-CoV-2 detection

Rapid, reliable, large-scale detection of the newly emerged coronavirus SARS-CoV-2 is crucial for controlling the COVID‑19 pandemic. An effective SARS-CoV-2 detection method must be able to detect just a few copies of the virus in a streamlined protocol that simplifies sample preparation and handling to reduce the risk of contamination, delivers fast, precise, and reproducible results, and can process a large number of samples per day.

Real-time quantitative PCR (RT-qPCR), a powerful tool for gene expression analysis, forms the basis for a wide variety of viral detection methods. A number of specialized RT-qPCR-based detection methods are available that can be used to develop effective SARS-CoV-2 detection protocols. One-step RT-qPCR allows efficient, specific cDNA synthesis and amplification in a single tube, reducing sample handling time and minimizing the risk of contamination. Direct RT-qPCR permits viral detection directly from crude biological samples, eliminating the need for time-consuming RNA purification while maintaining accuracy and sensitivity. Automated, high-throughput SARS-CoV-2 detection enables large-scale sample processing with precision, reproducibility, and speed.


Sensitive, reliable viral detection with one-step RT-qPCR

When SARS-CoV-2 was first identified, Takara Bio’s one-step PrimeScript RT-qPCR technology was used to develop the first SARS-CoV-2 screening test (Zhu et al. 2020). These high-quality reagents, manufactured under ISO 13485:2016 conditions, have been utilized in multiple Emergency Use Authorizations issued by the Federal Drug Administration (FDA) to Ipsum Diagnostics, LLC, Hackensack University Medical Center, and LifeHope Laboratory for COVID-19 testing. Our RT-qPCR kits, which include the One Step PrimeScript III RT-qPCR Mix, the most sensitive one-step RT-qPCR master mix on the market, continue to be used worldwide with established primers for sensitive detection of SARS-CoV-2 down to five viral copies (Figure 1).

Reliable detection of SARS-CoV-2 with our one-step RT-qPCR kits

Figure 1. Reliable detection of SARS-CoV-2 at 5, 50, 500, 5,000, and 50,000 copies with our one-step RT-qPCR kits. These data were generated by Takara Bio using publicly available protocols from the CDC. These data have not been validated by the CDC.

In addition, researchers in two independent studies confirmed the superior performance of One-Step PrimeScript III RT-qPCR Mix compared to other one-step RT-qPCR solutions:

In an April, 2020 bioRxiv paper, Brown et al., from the Great Ormond Street Hospital (NHS, London, UK), compared One Step PrimeScript III RT-qPCR Mix to three leading competitors in order to test compatibility and sensitivity for SARS-CoV-2 detection. The authors showed that the lower limit of detection for this kit is 10 to 100 times greater than the competitors' solutions. They also demonstrated that the increased sensitivity of PrimeScript III one-step RT-qPCR reduces the number of false negatives. This study also compared the overall reaction speed for all the mixes and demonstrated that the PrimeScript III One Step RT-qPCR Mix outperforms other one-step RT-qPCR solutions in both sensitivity and speed.

In a July, 2020 medRxiv paper, Reijns et al. from the University of Edinburgh, also found that One-Step PrimeScript III RT-qPCR Mix demonstrated superior sensitivity when performing a multiplex reaction on patient samples. Critically, they report that this mix was able to detect viral RNA in samples that were previously found negative using a competitor kit. Moreover, they reported that the assay developed using the Takara Bio mix was 1/10th the price of the other kits. These data reinforce the exceptional performance and value of our mixes for SARS-CoV-2 detection.


Direct viral detection from crude biological samples—without RNA extraction

Our direct RT-qPCR mixes may be used directly with many types of biological samples, without the need for RNA purification, saving valuable time and resources. The Direct One-Step RT-qPCR Mix for SARS-CoV-2 is designed specifically for unpurified saliva samples, enabling accurate one-step RT-qPCR to detect the presence and amount of virus. The entire RNA extraction-free protocol takes less than one hour and can be used to process many samples at once (Figure 2, Panel A). Since this mix is based on the exceptionally sensitive One Step PrimeScript III RT-qPCR Mix, one can expect it to provide comparable sensitivity. The performance of Direct One-Step PrimeScript RT-qPCR Mix for SARS-CoV-2 with unpurified saliva samples was confirmed across a range of target RNA copy numbers (Figure 2, Panel B).

SARS-CoV-2 Saliva-swab test

Figure 2. The Direct One-Step PrimeScript RT-qPCR Mix for SARS-CoV-2 uses a simple 3-step protocol to detect SARS-CoV-2 in saliva samples across a range of target RNA copy numbers. Panel A. Our simple protocol takes less than an hour to complete. Panel B. Unpurified saliva samples spiked with synthetic RNA of the N1 and N2 regions of SARS-CoV-2 were processed according to the Direct One-Step PrimeScript RT-qPCR Mix for SARS-CoV-2 protocol, and positive detection was achieved for these targets as well as human RNase P (RP), the positive control.

Direct RT-qPCR from a variety of biological samples, such as viruses, bacteria, cultured cells, whole blood, and stool samples, is possible with the PrimeDirect Probe RT-qPCR Mix. Many protocol guidelines for SARS-CoV-2 detection have been developed using this technology. Lübke et al. described their simple SARS-CoV-2 detection protocol for respiratory samples using PrimeDirect Probe RT-qPCR Mix in a Journal of Clinical Virology paper.


SmartChip Real-Time PCR System

High-throughput viral detection via qPCR

The SmartChip Real-Time PCR System automates SARS-CoV-2 detection, allowing processing of a large number of samples with precision, reproducibility, and speed. It minimizes false positives and negatives, accurately captures transcripts down to single-digit copy numbers, reduces hands-on time, and maximizes the number of samples processed per day. This system is a complete solution that includes chips and reagents, allowing the processing of thousands of samples per day. Each well of the 5,184-well chip holds nanoliter-scale reaction volumes, eliminating a preamplification step for reduced variability and decreasing costs.


Webinar: High-throughput SARS-CoV-2 detection data using the SmartChip Real-Time PCR System

We have generated proof-of-concept data from assays run on the SmartChip Real-Time PCR System that indicate sensitive and reproducible detection of human RNase P and viral nucleocapsid genes. In addition, we have developed an in-house SmartChip protocol to detect SARS-CoV-2. Sign up to watch a webinar describing the full data set.


Webinar: Detecting and quantifying SARS-CoV-2 in wastewater using the SmartChip Real-Time PCR System

The SmartChip Real-Time PCR System has been used to detect and quantify antibiotic resistance genes and SARS-CoV-2 in wastewater, which can reveal important information about COVID-19 transmission, as described in a webinar by Dr. Windi Muziasari, cofounder and CEO of Resistomap. Sign up to watch the webinar and learn how.


References

  • CDI Enhanced COVID-19 Test EUA Summary, Hackensack University Medical Center (HUMC) Molecular Pathology Laboratory (April 15, 2020). Available at: https://www.fda.gov/media/137036/download
  • COV-19 IDx assay EUA Summary, Ipsum Diagnostics, LLC (April 1, 2020). Available at: https://www.fda.gov/media/136621/download
  • Brown, J. R. et al. Comparison of SARS-CoV2 N gene real-time RT-PCR targets and commercially available mastermixes. bioRxiv 2020.04.17.047118; doi: https://doi.org/10.1101/2020.04.17.047118
  • Lübke, N. et al. Extraction-free SARS-CoV-2 detection by rapid RT-qPCR universal for all primary respiratory materials. Clin. Virol. 130, 104579 (2020). Available at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7405857/
  • Reijns, M. AM. et al. A sensitive and affordable multiplex RT-qPCR assay for SARS-CoV-2 detection. medRxiv 2020.07.14.20154005; doi: https://doi.org/10.1101/2020.07.14.20154005
  • Zhu, N. et al. A novel coronavirus from patients with pneumonia in China, 2019. N. Engl. J. Med. NEJMoa2001017 (2020); doi: https://www.nejm.org/doi/full/10.1056/NEJMoa2001017
    • Zhu et al. supplementary appendix: RT-qPCR protocol and primers used 

Featured products

Cat. # Product Size Price License Quantity Details
638329 Direct One-Step RT-qPCR Mix for SARS-CoV-2 200 Rxns USD $359.00

Takara Bio’s Direct One-Step RT-qPCR Mix for SARS-CoV-2 allows the detection of SARS-CoV-2 in unpurified saliva samples, following pretreatment with the included Sample Prep Solution (not sold separately).

Cat. No. 638329 provides enough reagents for 200 25-µl reactions.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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RR600A One Step PrimeScript™ III RT-PCR Kit 200 Rxns USD $342.00

One Step PrimeScript III RT-qPCR Mix is a dedicated reagent for one-step real-time, probe-based RT-qPCR (using the 5’ nuclease method). This 2X premix does not freeze at its storage temperature of –20℃, so a reaction can be started simply by adding the template sample, primer, and a probe for detecting the desired target. The quick and simple protocol allows the reverse transcription and qPCR reactions to be performed in the same tube. The reverse transcription reaction uses the novel PrimeScript III RTase, which displays increased heat tolerance (up to 55℃) while maintaining the specificity and extensibility of PrimeScript RTase. This allows cDNA synthesis from RNA with a more complex secondary structure. After cDNA synthesis, TaKaRa Taq HS performs highly specific and efficient PCR amplification, while the fluorescence emitted by the probe is detected in real-time. One Step PrimeScript III RT-qPCR Mix is also highly resistant to a wide variety of inhibitory substances in blood and soil, allowing stable one-step real-time RT-qPCR to be performed on a wide range of samples. This product can be used for various applications such as gene expression, RNA virus detection, etc.

Cat. # RR600A contains sufficient reagent for performing 200 reactions in a volume of 25 µl per reaction.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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RR600A: One Step PrimeScript III RT-PCR Kit

RR600A: One Step PrimeScript III RT-PCR Kit
RR064A One Step PrimeScript™ RT-PCR Kit (Perfect Real Time) 100 Rxns USD $337.00

The One Step PrimeScript RT-PCR Kit (Perfect Real Time) is designed for one step, real-time reverse transcription PCR (RT-PCR) using probe detection. With this kit, all RT-PCR steps can be performed in a single tube. Therefore, it is not necessary to add additional reagents during the reaction, minimizing the risk of contamination and simplifying the workflow. Amplified products are monitored in real time without need for gel electrophoresis after PCR. This kit is suitable for detection of small amounts of RNA. This kit uses PrimeScript Reverse Transcriptase, which allows efficient and rapid cDNA synthesis, and Takara Ex Taq HS, which enables highly efficient hot start PCR.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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RR064A: One Step PrimeScript RT-PCR Kit (Perfect Real Time)

RR064A: One Step PrimeScript RT-PCR Kit (Perfect Real Time)
RR650A PrimeDirect™ Probe RT-qPCR Mix 200 Rxns USD $340.00

PrimeDirect Probe RT-qPCR Mix is designed for one-step, real-time RT-PCR via probe detection (5'-nuclease method). This product is supplied as a 2X premix to facilitate the easy preparation of reaction mixtures. It requires only the addition of your primer, probe, and sample, and the RT-qPCR can be performed by simply adding your sample directly to the reaction mixture without intervening nucleic acid purification steps. Because this product is highly resistant to various PCR inhibitors in blood and soil, it can be used to directly detect pathogens, including viruses and bacteria, in various biological specimens and to directly analyze gene expression in cells.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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RR650A: PrimeDirect Probe RT-qPCR Mix

RR650A: PrimeDirect Probe RT-qPCR Mix
640022 SmartChip™ Real-Time PCR System Each Inquire for Quotation

License Statement

ID Number  
350 This product is protected by one or more of these U.S. Patents 7,622,296, 9,909,171, 9,132,427, 9,228,933 or 9,951,381, and corresponding foreign patents. Additional patents are pending. For further license information, please contact a Takara Bio USA licensing representative by email at licensing@takarabio.com.
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The SmartChip Real-Time PCR System offers high throughput genotyping and gene expression analysis in SmartChips which contain 5,184 individual nanowells. Samples and assays are dispensed into the SmartChip using the MutliSample NanoDispenser (MSND). Once the samples and assays have been dispensed, the SmartChip can be thermal cycled in the SmartChip Real-Time PCR Cycler.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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640022: SmartChip Real-Time PCR System

640022: SmartChip Real-Time PCR System

*You must be logged in to a Business Account in order to purchase these products online, since the purchase of these products may be restricted depending on your account type. Researchers at not-for-profit accounts receive a limited use license with their purchase of the product. Researchers at for-profit accounts must obtain a license prior to purchase. For details please contact licensing@takarabio.com.

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