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Pre-PCR: When Generating PCR-Compatible Samples Becomes Possible

Pre-PCR, qPCR, digital PCR

Peter Rådström’s research is about the developments of pre-PCR processing, a method which aims to generate PCR-compatible samples. Diagnostic PCR is often limited by components that interfere with the amplification of PCR inhibitors and pre-PCR can overcome PCR inhibition without impairing the detection limit, addressing all steps leading to PCR amplification, samples treatment and PCR chemistry. This lowers the need for pure extracts, leading to simplified analysis processes and improved detection limits.

We interviewed him about the research he is conducting at Lund University and the future advances that pre-PCR can bring to his field.

Research focus in pre-PCR processing

The main focus of the research is the bottleneck occurring along the process of PCR and how researchers can overcome it through the advanced technology that next-generation sequencing and digital PCR offers. In some ways, this research relates to a low-tech area, but it is something which is really needed particularly in sectors such as food, agriculture and forensics.

Nevertheless, there is not a huge need when it comes to the medical side. If it is sepsis, for example, there exists a necessity to detect something fast as it would be better to diagnose it before it is too late. Broadly speaking it is something needed for treatments. For a few years, we have been collaborating with different labs and have been examining all the different fields of PCR when it comes to diagnostics, putting all of our heads together to enhance the development of sample preparations.

Future developments in the field

In Sweden, there is a growing interest in developing methods based on the concept of pre-PCR processing, in which adjust the chemistry in PCR amplification. It’s suitable for the sample itself and also all the reagents in preparation of the sample, so you have something that is compatible all the way. In fact, it makes it possible to redefine the identity as well as the nature of inhibitors.

The novelty is that it makes it possible to streamline the chemistry in the entire workflow and start adjusting the chemistry reaction itself by using the right type of enzyme. By adding facilitators, you are merely helping and handling the inhibitors that were there from the start, applying the right type of buffer. There are a lot of possibilities today because there are so many different types of enzymes out in the market.

Furthermore, we know more about the mechanism of inhibitions occurring in the reaction and how we can bypass and change these. I think this represents an important driving force to develop further research in this area and validate new computational methods.

Peter Radstrom
Peter Rådström is Professor and Head of Applied Microbiology at Lund University, Sweden. Beside pre-PCR, he is also an expert in food microbiology, food safety and water quality. He participated in the 4th qPCR & Digital PCR Congress.


What are the other novelties in qPCR and digital PCR? Join the 3rd qPCR & Digital PCR Congress: USA and find out more.

 

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