ANINA: Molecular Accuracy Without the Laboratory.

How a pond in Salt Lake City led Seek Labs to replace thermal control with spatial control

By Jared Bauer, CEO and Co-Founder, Seek Labs

Recently, we announced the publication of research conducted by our scientists at Seek Labs in Nature Communications. The paper introduces Annexing Isothermal Nucleotide Amplification, or ANINA™, a new amplification chemistry designed to deliver the sensitivity and specificity of molecular testing without the equipment and infrastructure traditionally required to perform it.

The published results are significant. ANINA amplified genetic material at 25°C within 30 minutes. It detected extremely low concentrations of viral DNA in complex biological backgrounds. It enabled a complete, equipment-free result in under 45 minutes. And in its real-time format, it matched qPCR’s quantitative performance while producing answers substantially faster. Read the full ANINA publication in Nature Communications.

But the numbers alone do not explain why we built it or how hard we pushed to make it this simple.

The Pond Set the Standard

Nearly a decade ago, our scientists carried one of our early molecular tests out of the laboratory and took it to a pond in Salt Lake City.

We didn’t go to the pond to conduct an experiment. We went to evaluate the environment. We wanted a place without a clean bench, a calibrated instrument, a refrigerator, or a plug in the wall. Standing beside the pond forced a hard question: could we redesign our test so that molecular accuracy no longer required a laboratory to produce it?

Not just a portable version of a laboratory or a smaller instrument housed inside a cartridge. A molecular test designed from the chemistry up to work without thermal cycling, cold chain, power, microfluidic chips, or trained operators.

The setting made every hidden assumption visible. What did the test require from its surroundings? What did it require from the people using it? And what would we have to change for laboratory-quality molecular accuracy to reach the places a laboratory cannot go?

We did not solve those problems that day.

We failed. Then we failed again. Over years, our team tried different approaches, learned from what did not work, and kept engineering the chemistry’s dependence on laboratory infrastructure out of the reaction without giving up the sensitivity or specificity that makes a molecular result worth trusting.

That work has now been published in Nature Communications.

What the paper shows is that the standard we set beside that pond was not unrealistic. Molecular testing does not have to stay where the laboratory is. Analytical performance does not have to depend on the infrastructure that has historically carried it. And accuracy at the point of need does not have to be traded for the conditions in which the answer is needed.

The pond gave us that standard; ANINA is the science that met it.

From Salt Lake City to Vietnam

White Spot Syndrome Virus is the use case that forced our team to confront what “without the laboratory” actually demands.

WSSV can move through a shrimp population with devastating speed. Molecular tests can detect it early, but those tests typically depend on centralized laboratories: samples shipped on ice, processed on a benchtop instrument by a trained operator, results returned days later. By the time results are available, the pond is already lost.

Dr. Annie Roy, Vice President of Diagnostic Research at Seek Labs, used this problem to push our team past incremental thinking. After the early pond exercise, the team eventually took the technology to a shrimp facility in Vietnam and evaluated it in the kind of real-world environment the chemistry had been designed for: ambient tropical temperatures, no cold chain, no benchtop instrument, and a result the farmer could act on before the virus moved through the shrimp population. The conditions were no longer hypothetical. They were the conditions the chemistry had been built to handle.

That progression—from a pond in Salt Lake City, to a shrimp facility in Vietnam, to the controlled oral-exposure study reported in the paper—reflects how ANINA was developed. We did not begin with an existing laboratory method and ask how to make the equipment smaller. We began with the environment where the answer was needed and asked what the chemistry would have to become in order to work there.

As Dr. Roy put it: “This is a chemistry advance, not a smaller box for an existing test.”

The publication does more than report a promising test result. It describes a new mechanism for molecular amplification and demonstrates that removing the laboratory did not require us to give up analytical performance.

The Problem with Traditional Laboratory-based Testing

PCR achieves high specificity partly through thermal cycling. Each temperature change helps control when primers can bind and amplification can proceed. But that performance depends on infrastructure the chemistry itself cannot supply: thermocyclers, electricity, cold chain, trained operators, and the laboratory around them.

Earlier isothermal methods removed thermal cycling but introduced different constraints. LAMP typically operates at 60°C to 65°C and uses four to six primers. RPA operates at lower temperatures but generally requires long primers and can produce background amplification. Other approaches add enzymes or separate reactions to improve specificity and detection.

That is the challenge our team set out to solve. It was not enough to remove the thermocycler. We had to replace, inside the chemistry, the control the thermocycler used to provide.

How ANINA Replaces Thermal Control with Spatial Control

ANINA does not simply remove heat. It replaces thermal control with spatial control.

At the center of the technology is the Annexing Probe. A recombinase—an enzyme that helps locate matching genetic sequences—guides the probe to the correct target and opens the DNA locally. That opening recruits a short adjacent primer into the right position to begin amplification.

The primer is not left to find and engage the target on its own. The probe organizes that interaction spatially. This is what differentiates ANINA from earlier approaches. The Annexing Probe is simultaneously the regulator, the specificity gate, and the detection handle. That distinction matters. A molecular amplification method must do more than generate signal. It must generate the right signal. A chemistry that runs at ambient temperature but amplifies the wrong sequence is not useful.

The paper provides direct evidence that the Annexing Probe controls the reaction. Identical primer sets performed poorly or failed without the probe. Moving the probe farther from the adjacent primer reduced amplification even when primer length remained unchanged. A single mismatch at the primer’s critical 3′ end substantially reduced amplification, while two mismatches eliminated detectable amplification.

Those findings show that ANINA preserved stringent sequence discrimination at ambient temperature. We did not trade specificity for the operating conditions we wanted.

The Role of the Annexing Probe

The Annexing Probe also remains associated with the amplified product, which allows the result to be read directly on a lateral-flow strip. The same molecule that helps regulate amplification also enables detection without a probe-cleavage enzyme, a CRISPR/Cas reaction, or a separate detection step.

That matters because every additional enzyme, reaction, transfer, or instrument reintroduces something the laboratory used to supply. ANINA addresses both sides of the isothermal challenge at once: it enables amplification at ambient temperature while preserving the specificity a meaningful molecular result requires.

It also expands where assays can be designed. Conventional RPA generally requires primers approximately 30 to 36 nucleotides long and typically operates between 30 °C and 42 °C. ANINA supported probe-adjacent primers as short as 10 nucleotides and downstream primers as short as 15 nucleotides while operating at ambient temperature. This is a different mechanism for controlling molecular amplification, not a repackaging of an existing one.

The Science Behind the Standard

The pond did not give us the answer. It gave us the standard. It forced us to ask what molecular testing would have to become if we removed the laboratory rather than shrinking it.

ANINA is our answer to that question. It replaces thermal control with spatial control. It uses the Annexing Probe to regulate amplification, preserve specificity, and enable direct detection. And it does so at ambient temperature, without the infrastructure that molecular accuracy has historically required. A new mechanism, though, only matters if the results hold up where the laboratory cannot follow.

In the next article in this series, I will explain what ANINA demonstrated in both equipment-free settings and standard laboratories, how those results connect to SeekIt™, and what still has to be proven before this technology reaches its full potential.