As we are still living in a pandemic world, Covid19 remains a lingering issue – not only for the general public but also for the researchers and scientists working to eradicate the virus. But before the virus can be eradicated – it first must be detected.
One popular detection method in the scientific community, as many are aware is the RT-qPCR method, otherwise known as a Quantitative Reverse Transcriptase Polymerase Chain Reaction.
According to the PubMed article, “Nucleic Acid Testing of SARS-CoV-2”, “the polymerase chain reaction (PCR) – based method including reverse transcription-polymerase chain reaction (RT-PCR) is the most widely used assay for the detection of SARS-CoV-2 RNA”.
But before we get too far, RT-PCR is the third piece to the Covid-19 detection puzzle so to speak. The first two “pieces”, include sample collection, followed by RNA extraction, and then what shall be discussed today – RT-PCR.
[To learn more about the first two “puzzle pieces”, check out these blogs for more information]
To start, let us first discuss how the process works:
- A sample is collected
- Once a sample is collected, RNA is extracted, and a Complementary DNA (cDNA) strand is created.
- Next, DNA polymerase, is added to the cDNA to create a double-strand à which then feeds into the *PCR amplification process. [*The PCR amplification process is done using fluorescent probes, which helps machines to identify the section of interest]
- In other words, the cDNA = Reverse Transcriptase, and the process of amplification by PCR collectively forms the RT-PCR (Reverse Transcriptase Polymerase Chain Reaction).
- Following the amplification process, the next step is to now add the SARS-COV-2 Primers and Probes to the RT-qPCR reaction, as well as the patient RNA sample.
- Once they are added, a COVID-19 infection can be finally be detected (if the virus is present of course)
While the RT-PCR Process might be nothing new for you as a researcher, there are also a few interesting alternative methods that are out in the scientific community that are also capable of detecting viruses.
Chart Source: Nucleic Acid Testing of SARS-CoV-2 (nih.gov)
As the chart shows, there are quite a few alternative methods for detecting viruses -LAMP, RPA, SDA etc. However, in regards to COVID-19 the PCR detection method is the most widely used – and for good reason.
According to frontiersin.org (in the article “A Basic Guide to Real Time PCR in Microbial Diagnostics), “the main advantages of qPCR are that it provides fast and high-throughput detection and quantification of target DNA sequences in different matrices…moreover, qPCR is safer in terms of avoiding cross contaminations because no further manipulation with sample is required after the amplification. Other advantages of qPCR include a wide range for quantification and the multiplexing of amplification of several targets into a single reaction.”
All of these things are ideal when detecting a virus because any contamination – even a microscopic one can throw an entire sample off – not only ruining your research but also your day.
As noted in the PubMed article, “Nucleic Acid Testing of SARS-CoV-2”, “most countries use the RT-qPCR assay as a primary method for diagnostics. Though alternative methods are available, their sensitivity, specificity, and costs are not comparable to RT-qPCR…”
So, as you can see, there are many benefits to using qPCR over the other alternative methods that exist in the scientific community. Ultimately when it comes down to getting a reliable and trustworthy SARS-COV detection, qPCR has got your back.
Now you might be wondering what you should use to complete your qPCR process to make a [potential] SARS-COV detection?
Well, you are in luck, because IBI just happens to carry the product for such a process.
In particular, IBI Scientific carries UniPLUS RT-qPCR Master Mix, which is a universal Master Mix that contains reagents and master mixes that are necessary to accurately and efficiently synthesize and amplify cDNA from RNA – which is perfect for COVID detection.
In addition, our Master Mix [Kit] contains an intercalating dye, and supplemental ROX reference dye (which is very useful due to its fluorescent quality - which makes it easy for machines to pick up).
Alongside the florescent quality of the ROX dyes in our kit, the ROX dyes enable use on a wide variety of instruments – which makes our kit usable for numerous labs – regardless of the available apparatuses.
Furthermore, another benefit to our Master Mix, is that its enzyme and buffer system has been optimized to support both 1-step and 2-step RT-qPCR protocols which gives you – the researcher the ability to choose which protocol method best suits you.
In addition, as many in the scientific community are aware, coming to a SARs-COV-2 diagnosis doesn’t always go as smoothly one might hope. There might be sample collection difficulty or the lab equipment might be slow – but in any case, having a trustworthy process can give much-needed peace of mind.
That’s why using a qPCR process is so beneficial – because it sets you and your lab up for success. As frontiersin.org (in “A Basic Guide to Real Time PCR…”), notes “qPCR is…a powerful tool…” and it will continue to be as we are still fighting the ever-present threat of COVID-19.
Here at IBI Scientific, we care strongly about the needs and potential concerns of our customers and scientific community. We also support the increasing number of research centers that are working vigorously to fight the battle against COVID-19 and its numerous variants.
May we end this pandemic together.
To buy any of our various life science research approved products, click the following links:
Still, have lingering questions? Feel free to email us at firstname.lastname@example.org and we would be happy to help!
Written by: Katie Draves
Nucleic Acid Testing of SARS-CoV-2 - PubMed (nih.gov) Yoo HM, Kim IH, Kim S. Nucleic Acid Testing of SARS-CoV-2. Int J Mol Sci. 2021 Jun 7;22(11):6150. doi: 10.3390/ijms22116150. PMID: 34200331; PMCID: PMC8201071.
Grace Adams; A beginner’s guide to RT-PCR, qPCR and RT-qPCR. Biochem (Lond) 22 June 2020; 42 (3): 48–53. doi: https://doi.org/10.1042/BIO20200034
Kralik P and Ricchi M (2017) A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything. Front. Microbiol. 8:108. doi: 10.3389/fmicb.2017.00108