On-Site Viral RNA Detection In Wastewater With Paper And Wax Microfluidics

Schematic version of on-site wastewater analysis using the microfluidic strips (Credit: Yuwei Pan et al., Cell, 2024)

Wastewater sampling has become a popular way over the years to keep track of the health of a population, including human ones, as pathogens are often detectable in the effluence from toilets. Since most houses connected to the centralized sewer systems, this means that a few sampling sites suffice to keep tabs on which viruses are circulating in an area. While sampling this wastewater is easy, the actual RNA analysis using PCR (polymerase chain reaction) still has to be performed in laboratories, adding complex logistics. An approach for on-site analysis using microfluidics was tested out by [Yuwei Pan] et al., as recently published in Cell.

This particular approach uses RT-LAMP (reverse-transcription loop-mediated isothermal amplification) to increase the amount of genetic material, which has the significant benefit over PCR that it does not require multiple thermal cycles, instead being run at a constant temperature. The filter paper used as the basis has wax microchannels printed on it, which help to guide the filtered wastewater to the reaction chambers. This is in many ways reminiscent of the all too familiar linear flow self-tests (RAT: rapid antigen test) that have become one of the hallmarks of the SARS-CoV-2 pandemic.

What this paper microfluidic device adds is that it doesn’t merely contain antigens, but performs the lysis (i.e. breakdown of the virus particles), genetic material multiplication using RT-LAMP and subsequent presence detection of certain RNA sequences to ascertain the presence of specific viruses. Having been used in the field already since 2020 in the UK, the researchers envision this type of on-site analysis to be combined with a smartphone for instant recording and transmission to health authorities.

Some of the benefits of this approach would be lower cost, easier logistics and faster results compared to shipping wastewater samples to central laboratories.

Sequencing DNA For Metagenomics

If someone brought you an odd piece of electronic hardware and you wanted to identify it, you’d probably look for markings on the outside first. If that didn’t work out, you might look under the cover and read some markings on the board or key components. However, in a tough case, you might dump the firmware and try to guess what the device is or what it does by examining the code that makes it run. That’s kind of what [Ciro] did. Wanting to determine the bacteria in a water sample led to using relatively inexpensive DNA sequencing hardware to look at the DNA present in the samples. This would have been a huge undertaking for a well-funded lab just a few short years ago. Now it just takes a USB device and some software.

Of course, inexpensive is in the eye of the beholder. The micropore sequencer costs about $500 and has a one-time use consumable cost of about $500, although that’s enough to process about 10 human genomes. The technology depends on using a small pore only large enough to pass one strand of DNA at a time. Blocks of nucelotides cause different amounts of electrical current to flow through the pore.

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Defense Department Funds Wearables To Detect COVID-19

As many countries across the globe begin loosening their stay-at-home orders, we’re seeing government agencies and large companies prepare for the lasting effects of the pandemic. A recent solicitation from the United States Department of Defense (DoD) indicates they are investing $25 million into wearable devices that can detect early signs of COVID-19.

Based on a few details from the request for project proposals, it looks like the DoD is targeting mostly companies in this particular solicitation, but have left the door open for academic institutions as well. That makes intuitive sense. Companies can generally operate at a faster pace than most academic research labs. Given the urgency of the matter, faster turnarounds in technological development are imperative. Nonetheless, we have seen quite a bit of important COVID-19 work coming from academic research labs and we imagine that battling this pandemic will take all the brilliant minds we can muster together.

It’s good to see the DoD join the fight in what could be a lengthy battle with the coronavirus.

Please feel free to read through the request for project proposals for more details.

Hackaday Podcast 060: Counting Bees, DogBox Transmissions, And The Lowdown On Vents, BiPAP, And PCR

Hackaday editors Elliot Williams and Mike Szczys recount the past week in hardware hacking. There’s a new king of supercomputing and it’s everyone! Have you ever tried to count bees? Precision is just a cleverly threaded bolt away. And we dig into some of the technical details of the coronavirus response with a close look at PCR testing for the virus, and why ventilators are so difficult to build.

Take a look at the links below if you want to follow along, and as always tell us what you think about this episode in the comments!

Take a look at the links below if you want to follow along, and as always, tell us what you think about this episode in the comments!

Direct download (60 MB or so.)

Continue reading “Hackaday Podcast 060: Counting Bees, DogBox Transmissions, And The Lowdown On Vents, BiPAP, And PCR”

Hackaday Podcast 055: The Most Cyberpunk Synthesizer, Data In Your Cells, Bubbly In Your Printer, And The Dystopian Peepshow

Hackaday editors Mike Szczys and Elliot Williams discuss the many great hacks of the past week. Just in case you missed the fact that we’re living in the cyberpunk future, you can now pop off your prosthetic hand and jack directly into a synthesizer. The robot headed for Mars has a flying drone in its belly. Now they’re putting foaming agent in filament to make it light and flexible. And did you ever wonder why those pinouts were so jumbled?

Take a look at the links below if you want to follow along, and as always tell us what you think about this episode in the comments!

Take a look at the links below if you want to follow along, and as always, tell us what you think about this episode in the comments!

Direct download (60 MB or so.)

Continue reading “Hackaday Podcast 055: The Most Cyberpunk Synthesizer, Data In Your Cells, Bubbly In Your Printer, And The Dystopian Peepshow”

DNA Now Stands For Data And Knowledge Accumulation

Technology frequently looks at nature to make improvements in efficiency, and we may be nearing a new breakthrough in copying how nature stores data. Maybe some day your thumb drive will be your actual thumb. The entire works of Shakespeare could be stored in an infinite number of monkeys. DNA could become a data storage mechanism! With all the sensationalism surrounding this frontier, it seems like a dose of reality is in order.

The Potential for Greatness

The human genome, with 3 billion base pairs can store up to 750MB of data. In reality every cell has two sets of chromosomes, so nearly every human cell has 1.5GB of data shoved inside. You could pack 165 billion cells into the volume of a microSD card, which equates to 165 exobytes, and that’s if you keep all the overhead of the rest of the cell and not just the DNA. That’s without any kind of optimizing for data storage, too.

This kind of data density is far beyond our current digital storage capabilities. Storing nearly infinite data onto extremely small cells could change everything. Beyond the volume, there’s also the promise of longevity and replication, maintaining a permanent record that can’t get lost and is easily transferred (like medical records), and even an element of subterfuge or data transportation, as well as the ability to design self-replicating machines whose purpose is to disseminate information broadly.

So, where is the state of the art in DNA data storage? There’s plenty of promise, but does it actually work?

Continue reading “DNA Now Stands For Data And Knowledge Accumulation”

Put The Power Of PCR In Your Pocket With This Open-Source Thermal Cycler

When the first thermal cyclers for the polymerase chain reaction came out in the 1980s, they were as expensive as a market driven by grant money could make them. Things haven’t got much better over the years, largely shutting STEM classes and biohackers out of the PCR market. That may be about to change, though, if the €99.00 PocketPCR thermal cycler takes hold.

PCR amplifies DNA in a three-step process: denaturation, which melts double-stranded DNA into single strands; annealing, which lets small pieces of primer DNA bind to either side of the region of interest; and elongation, where the enzyme DNA polymerase zips along the single strands starting at the primer to replicate the DNA. The cycle repeats and copies of the original DNA accumulate exponentially. Like any thermal cycler, [Urs Gaudenz]’s PocketPCR automates those temperature shifts, using a combination of PCB-mounted heating elements and a cooling fan. The coils rapidly heat a reaction block up to the 99°C denaturation temperature, the fan brings that down to the 68°C needed for annealing, and then the temperature ramps back up to 72°C  for elongation with thermostable DNA polymerase. PID loops keep the reaction temperature precisely controlled. The whole thing is, as the name suggests, small enough to fit in a pocket, and can either be purchased in kit form or scratch-built from the build files on GitHub.

We applaud [Urs]’ efforts to get the power of PCR into the hands of citizen scientists. Quick and dirty thermal cyclers are one thing, but Pocket PCR has a great fit and finish that makes it more accessible.

Thanks to [Abe Tusk] for the tip.