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The recent emergence of the coronavirus SARS-CoV-2 (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 COVID-19 down to five viral copies (Figure 1).
Viral detection directly from biological samples—no RNA purification
Detection of inactivated influenza A virus H1N1
PrimeDirect Probe RT-qPCR Mix may be used directly with many types of biological samples, without the need for RNA purification, saving valuable time and resources (Figure 2). We performed H1N1 detection directly on biological samples using PrimeDirect Probe RT-qPCR Mix, following the protocol shown below. Inactivated influenza A virus was spiked into mouth swab samples, nasal swab samples, and saliva samples at a concentration of 2 x 100–102 copies/µl.
Soak the sampled mouth swab into 200 µl PBS buffer, shake for 5 seconds, and then add 1 µl of supernatant to reaction.
Nasal cavity swab
Soak the sampled nasal swab in 200 µl PBS buffer, shake for 5 seconds, and then add 1 µl of supernatant to reaction.
Directly add 1 µl of supernatant to reaction.
PrimeDirect Probe RT-qPCR Mix (2X)
Forward Primer (10 µM)
Reverse Primer (10 µM)
qPCR probe (10 µM)
Influenza A virus H1N1 (2 x 100–102 copies/µl)
RNase Free H2O
Figure 2. PrimeDirect Probe RT-qPCR Mix enabled the detection of viral RNA directly from biological samples. The detection sensitivity from nasal cavity swab samples and saliva samples was 20 copies/μl, and the sensitivity from mouth swab samples was 2 copies/μl.
Compatibility with primer/probe sets from published COVID-19 detection protocols
We tested qPCR detection of COVID-19 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 COVID-19 detection (Zhu et al. 2019).
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Citations for COVID-19 detection using Takara Bio one-step RT-PCR kit
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.