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COVID-19 research

Real-time qPCR for COVID-19 research

The recent emergence of the coronavirus SARS-CoV-2 (the virus that causes COVID-19) has led to a need for rapid and reliable detection. Takara Bio offers solutions to support researchers and clinicians studying this novel coronavirus, such as viral RNA purification kits, NGS kits, and RT-qPCR kits that can be used with established primers for sensitive detection of SARS-CoV-2 down to five viral copies (Figure 1).

Takara Bio reagents in FDA Emergency Use Authorizations

Our high-quality reagents, manufactured under ISO 13485:2016 conditions, have been utilized in multiple Emergency Use Authorizations issued by the Federal Drug Administration (FDA) for COVID-19 testing:

COVID-19 detection using PrimeDirect RT-qPCR

Publications: highly sensitive, fast SARS-CoV-2 detection with PrimeScript III one-step RT-qPCR

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 QuantiFast Multiplex PCR +R master mix (Qiagen), TaqMan Fast Virus 1-Step Master Mix (Thermo), and TaqPath One Step RT-qPCR Master Mix (Thermo) in order to test compatibility and sensitivity for SARS-CoV-2 detection.

In the initial experiment to test the limit of detection, One Step PrimeScript III RT-qPCR Mix showed superior sensitivity by consistently detecting 1 viral copy/µl with all three primer/probe assays. The authors showed that the lower limit of detection for this kit is 10 to 100 times greater than the competitors' solutions.

Next, the authors tested 74 real-life samples from nose and/or throat swabs from patients with suspected cases of SARS-CoV-2 infection. They compared PrimeScript III mix to the Quantifast multiplex kit (Qiagen). Of the 74 swab samples tested, 67/74 showed concordant results between both master mixes. However, among the discordant results, PrimeScript III mix could detect 7 patient samples with low RNA sample inputs (median Ct = 40.9 [range 38.5–41.9]), whereas these samples were undetected by Quantifast, showing that the increased sensitivity of PrimeScript III one-step RT-qPCR reduces the number of false negatives.

Furthermore, the authors compared the overall reaction speed for all mixes. The PrimeScript III one-step RT-qPCR experiment can be run in less than 1 hour (33% faster than Quantifast Multiplex PCR +R master mix), meaning even faster detection of SARS-CoV-2 infection. Faster reaction speeds will help increase the throughput for COVID-19 diagnostic labs.

Brown et al. conclusively showed that the PrimeScript III One Step RT-qPCR Mix outperforms other one-step RT-qPCR solutions in both sensitivity and speed. In a time where screening for SARS-CoV-2 infection is of utmost importance, the efficiency and rapidity of One Step PrimeScript III RT-qPCR Mix plays a critical role in the optimal management and control of this pandemic.

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 positive samples that were previously found negative using the TaqPath COVID-19 Combo 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.

Compatibility with primer/probe sets from published SARS-CoV-2 detection protocols

We tested qPCR detection of SARS-CoV-2 using the One Step PrimeScript III RT-PCR Kit and PrimeDirect Probe RT-qPCR Mix in publicly available protocols with primer sets from the US and China CDCs, WHO, and Japan NIID. Sensitivity of the assay varies depending on the nature of the template. The template used in the Japan NIID protocol was provided by the NIID, and the templates used in the other protocols were synthesized RNA prepared by Takara Bio. You can download our results by clicking on the PDF thumbnail images below.

The China CDC has also published a paper using our kits for SARS-CoV-2 detection (Zhu et al. 2019).

We have bulk formats of our one-step RT-qPCR kits. If you are interested, please contact us with your inquiry.

Explore industry-leading products that can advance your COVID-19 research:

Cat. #	Product	Size	Price
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. Documents Components Image Data Back RR600A: One Step PrimeScript III RT-PCR Kit
RR600B	One Step PrimeScript™ III RT-PCR Kit	1,000 Rxns	USD $1538.00
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. # RR600B contains five of Cat. # RR600A. Please refer to Cat. # RR600A for complete product documentation and resources. 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.
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. Documents Components Image Data Resources Back RR064A: One Step PrimeScript RT-PCR Kit (Perfect Real Time)
RR064B	One Step PrimeScript™ RT-PCR Kit (Perfect Real Time)	500 Rxns	USD $1336.00
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. Cat. # RR064B contains 5 of Cat. # RR064A. Please refer to Cat. # RR064A for complete product documentation and resources. 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.

More Information

Citations for SARS-CoV-2 detection using Takara Bio RT-PCR kits

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Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

Save resources and increase sample throughput for COVID-19 detection

The SmartChip Real-Time PCR System is a complete, high-throughput solution with unrivaled flexibility in assay and sample formats. This system supports:

100-nl reaction volumes that decrease costs up to 200X vs. traditional plates
5,184 wells for processing up to 384 samples per chip
Automated dispensing that reduces hands-on time
14 different dispense patterns that enable rapid switching, adding, or removing assays and replicates

Learn more

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