Avoiding the itch with digital PCR

05/31/23 • 36 min

Parasites may bet a bad rap overall, but they play a vital role in healthy ecosystems. In this episode, we focus on the role parasites play in freshwater ecosystems. Specifically, we’re talking about the role of avian schistosomes, a very interesting parasite that infects waterfowl, but that also uses snails as a host in its larval stage. Larvae also infect humans to cause what’s know as swimmer’s itch.

To guide this conversation we have Dr. Patrick Hanington, associate professor in the School of Public Health at the University of Alberta. As a self-described parasitologist and immunologist he and his team focus on developing multiplexed PCR-based tests to detect freshwater parasites, including avian schistosomes. Their work benefits locals in his area by monitoring pubic and recreational waters for swimmer’s itch outbreaks, but their work also serves as a model for informing human schistosome research, where Schistosomiasis is the second most prevalent disease worldwide, behind malaria.

In our conversation with Patrick we learn about how they design their assays, why they’re increasingly using dPCR instead of qPCR. Beyond the technical work, we get into how Patrick’s career path developed, how what he loves most about his job has changed and evolved over time, his lessons learned in the lab, and how his research and hobbies have blended over time. And because it’s Absolute Gene-ius, you know we keep it fun with some unexpected movie references and a bit of discussion about how science is represented in television and film.

Visit the Absolute Gene-ius page to learn more about the guest, the hosts, and the Applied Biosystems QuantStudio Absolute Q Digital PCR System.

Visit the Absolute Gene-ius page to learn more about the guest, the hosts, and the Applied Biosystems QuantStudio Absolute Q Digital PCR System.

Previous Episode

Insights from a PCR Whisperer

Visit the Absolute Gene-ius page to learn more about the guest, the hosts, and the Applied Biosystems QuantStudio Absolute Q Digital PCR System.

The details of what make digital PCR (dPCR) different from real-time, or quantitative PCR (qPCR) are relatively simple but not always explained very well. Likewise, it’s not always clear which use cases are a good fit for dPCR, and which others simply don’t require the power of dPCR. The power of digital PCR is real, if you understand it.

In this episode we enlist Marcia Slater, a self-described “PCR guru” to explain digital PCR and its power. She covers the basic differences between dPCR and qPCR and then delves into the details of where dPCR derives its power and where it shines. With over 20 years’ experience in helping customers troubleshoot PCR, Marcia makes is easy to understand key terms and concepts related to dPCR, including:

Sub-reactions
Poisson statistics
Statistical power and confidence intervals
Controls and false negatives vs. true negatives
Dead volume
Dynamic range
Multiplexing

Marcia also covers some great examples of where the absolute quantification of dPCR is a great fit and how it’s even used to qualify and quantify standards for qPCR. Multiplexing and how its used to do molecular integrity evaluations for gene therapy applications is also discussed.

As always with the Gene-ius series, you’ll also get to learn about more than Marcia’s science chops. We learn about her unlikely career path from growing up on a livestock farm to her storied role in helping produce “data so beautiful it should be framed.” We even get into her rediscovered love of raising animals, including her beloved panda alpaca with a name you cannot forget!

Next Episode

Raising the fish that spawn our insights

The use of model organisms in science dates to ancient Greece and represents an important way humans have progressed our collective understanding of biology and disease. We’ve probably all heard of using mice, rats, or even Drosophila melanogaster (i.e., fruit flies) in the lab, but zebrafish have surged in their use as a model organism. Zebrafish are small, more cost effective to maintain, have a fast generational lifecycle, and have clear embryos that enable direct observation of their development. As is the case with any model system used across the globe, standards matter and help make results transferrable to other studies. This is where our guests come into the picture.

For this unique episode we’re joined by both Corbin Schuster and Zoltan Varga of Zebrafish International Resource Center (ZIRC) at the University of Oregon. In their roles, they help raise and maintain over 12,000 genetically unique zebrafish lines for use in studies across the globe. We learn about the health monitoring they conduct on a regular basis to maintain their colony and support their own and partner studies using their zebrafish. The conversation touches on:

Environmental DNA (eDNA) methods they employ
Assay development and method selection based on their need for sensitivity and specificity
The use of qPCR and dPCR in their work
Studies and monitoring of parasites and pathogens that affect their colony
How zebrafish have helped shift research from forward genetic studies to reverse genetic studies

As always, you’ll also get to know our guests on a more personal level. We learn about how they each got into this line of work, what they love about it, and how they both value human relationships and helping their communities. Through the audio alone you’ll get a sense of the positive, kind, and collaborative attitudes that have helped them both be successful in their careers.

Visit the Absolute Gene-ius page to learn more about the guests, the hosts, and the Applied Biosystems QuantStudio Absolute Q Digital PCR System.