FORTIFIEDx: Bringing true sample-to-result diagnostics to the point of care

We speak with a few project partners about the novel technologies behind and how it could help bring reliable testing closer to the patient.

Healthcare systems around the world increasingly depend on rapid diagnostics. Whether identifying infectious diseases early, preventing the spread of outbreaks, or reaching populations far from clinical laboratories, the ability to test quickly and reliably at the point of care has become one of modern medicine’s most pressing needs.

Yet despite decades of research and development, most point-of-care diagnostic technologies still fall short of delivering a fully integrated solution. Samples often need to be transferred, processed externally, or handled by trained personnel before a result can be obtained. In practice, the journey from sample to result frequently remains fragmented, costly and difficult to deploy outside laboratory environments.

The FORTIFIEDx project aims to change that. By combining novel sampling technologies, advanced microfluidics and integrated bioassays within a single disposable patch, the project is working towards what researchers describe as a true sample-to-result diagnostic system – one capable of performing the entire testing process autonomously at the point of care.

This innovation is driven by a clear objective: to overcome many of the practical barriers that still limit point-of-care testing today. “The idea is that we would like to develop a fully integrated disposable diagnostic patch based on microfluidics, to deliver a true sample-to-result test at the point of care,” explains Dragana Spasic, Research Manager in the Biosensors Group at KU Leuven. “Because this sample-to-result concept is still mostly missing when we talk about true point-of-care solutions.”

The FORTIFIEDx patch is designed to combine several processes that are usually carried out separately. Sampling, fluid handling, biochemical analysis and result generation are integrated into a single device intended to operate autonomously once activated.

“The idea behind this is to eliminate as much as possible the manual sample handling, external instrumentation and dependency on laboratories,” Spasic says. “All of these aspects are very important if we’re talking about point-of-care devices.”

Decentralised diagnostics
The motivation behind FORTIFIEDx reflects broader shifts in global healthcare. Increasingly, clinicians and public health systems are looking for ways to move diagnostics closer to patients. Part of that shift stems from the continued global burden of infectious diseases, including both established conditions and emerging pathogens. Another factor is the growing strain on laboratory infrastructure, highlighted starkly during the COVID-19 pandemic.

“While the demand for more decentralised care and better point-of-care solutions is clearly increasing, the technologies capable of delivering fast, reliable diagnostics directly at the point of care are still not fully there,” Spasic explains. “At the same time, the need for them is becoming more urgent.”

The urgency of that challenge is reinforced by a rapidly changing global landscape of infectious disease. Increased travel, environmental change and globalisation all contribute to the spread of pathogens across regions, placing greater pressure on healthcare systems to diagnose infections quickly and close to the patient.

For Dorien Van den Bossche, a microbiologist at the Institute of Tropical Medicine in Antwerp, these broader pressures are reflected in the ongoing challenges of diagnosing and controlling infectious diseases such as sexually transmitted infections like syphilis and HIV – diseases where timely and accessible testing remains critical.

“For syphilis, the latest numbers are certainly increasing,” she says. “So there is a need for reaching underserved populations, for example, to really bring testing towards the people that need the test.”

HIV presents similar challenges. While global trends show improvements in diagnosis and treatment, access to testing remains uneven. “UNAIDS has set a target to diagnose 95 per cent of people living with HIV by 2030,” Van den Bossche notes. “But we already struggle with reaching those numbers, so we really see that there is a need to expand testing services.”

Beyond these diseases, the diagnostic platform being developed in the project is also designed to address needs linked to emerging outbreaks, including viral haemorrhagic fevers such as Ebola and Lassa. “With the latest Ebola outbreaks, for example, we also noticed that there is a need to do safer testing at the point of care,” she adds. “And there the FORTIFIEDx patch would really help, because you don’t need a blood transfer anymore.”

Safer testing is only part of the challenge. In many infectious diseases, the ability to diagnose cases quickly and close to the patient is equally critical. Early diagnosis not only improves patient outcomes but also helps limit transmission. As Van den Bossche points out, this is a common challenge across many infectious diseases. “The earlier you can get a diagnosis, the more you reduce transmission and prevent further spread.”

Sample collection
Meeting these challenges requires more than incremental improvements to existing diagnostic tools. FORTIFIEDx approaches the problem by rethinking how point-of-care testing can be carried out, from the moment a sample is taken through to the delivery of a result. One of the project’s a few innovations in this sense lies in the way biological samples are collected. Instead of relying on traditional blood draws or finger-prick tests, the project integrates microneedle technology capable of extracting tiny quantities of interstitial fluid from the upper layers of the skin.

Developed by researchers at the Tyndall National Institute in Ireland, the microneedles offer a minimally invasive alternative to conventional sampling methods. “We’re looking at microneedle technologies to remove tiny amounts of fluid from the upper skin layers,” explains Conor O’Mahony, Principal Scientist at Tyndall. “This is in contrast to most diagnostic systems today, which rely on blood drawn from the finger.”

For many patients, blood sampling is uncomfortable and sometimes a barrier to testing altogether. “No one likes taking blood,” O’Mahony says. “It’s painful, it’s inconvenient, it’s messy.”

Microneedles operate on a different principle. Typically, around half a millimetre long, they penetrate only the uppermost skin layers without reaching nerve endings or blood vessels. “Because of that length, they don’t strike the blood capillaries and they don’t strike the nerve endings,” he explains. “So the use is both painless and bloodless to the user.”

The microneedles draw interstitial fluid, the liquid that surrounds cells within tissues. While less commonly sampled than blood, this fluid contains many of the same biological markers used for diagnostic testing. “That fluid is full of biomarkers,” O’Mahony says. “Almost everything that you find in blood is also present in interstitial fluid.”

The approach also addresses practical and social challenges associated with conventional needles. Around ten per cent of the population experiences severe needle phobia, a condition that can deter individuals from seeking medical care. “We call it trypanophobia,” he explains. “And that prevents quite a number of people from taking medication, from undergoing diagnostic procedures and even from going to the doctor in the first place.”

In addition, microneedles reduce the risk of needle reuse, an important safety issue in some parts of the world. “There’s an enormous trade in used needles in many developing countries,” O’Mahony says. “This technology eliminates that risk.”

The microfluidic engine
Once the sample has been collected, the challenge becomes processing it automatically within the device. This is where FORTIFIEDx’s novel microfluidic architecture comes into play. The system builds on iSiMPLE (infusion Self-powered Imbibing Microfluidic Pump by Liquid Encapsulation), a microfluidic platform developed by the KU Leuven Biosensors Group. The technology enables passive, finger-activated liquid handling without the need for external pumps or power sources, using pressure differences to move and control tiny volumes of fluid within the device. In the case of FORTIFIEDx, this enables complex biochemical reactions to be carried out on a compact chip in the patch.

“The big innovation is that we integrate everything into a standalone, self-powered device,” explains Jeroen Lammertyn, who leads the Biosensors Group coordinating the project.

“Inside the patch, fluids move through networks of channels roughly the width 8 to 10 times larger than that of a human hair,” Lammertyn explains. “These channels allow the system to measure, mix and process tiny volumes of liquid with remarkable precision.”

Because each patient sample can vary, the system must also control exactly how much fluid enters the reaction. “You have to take a metered sample,” he says, “and that sample may need to be diluted or combined with reagents.”

In traditional laboratory systems, these steps require pumps, instrumentation and trained operators. FORTIFIEDx replaces that complexity with a self-powered mechanism driven by capillary forces. “We use capillary paper as a kind of engine,” he says. “It creates pressure differences that allow us to move liquids through the chip and perform all the necessary fluid manipulations.”

This approach avoids one of the limitations of many existing rapid tests, which rely on lateral flow strips where samples are drawn through porous materials. “In lateral flow systems, molecules you are interested in can stick to the paper,” Lammertyn explains. “In our case, the sample itself moves through channels, while the paper only provides the driving force.”

The result is a level of control over fluid handling that is rarely achieved in portable diagnostic devices. “We can do very complex liquid manipulations on just two layers of plastic bound with double-sided tape,” he says.

Turning samples into results
Once the fluid is processed inside the microfluidic network, the system performs the biological analysis required to detect specific diseases. For sexually transmitted infections, the project integrates lateral flow assays capable of detecting both HIV and syphilis. Importantly, the syphilis component includes functionality not typically available in standard rapid tests.

“What is new compared to current self-tests is that we include a component that allows us to determine the activity of the disease,” Van den Bossche explains. “Current tests usually only detect whether someone has been exposed to syphilis in the past, but they cannot distinguish between active disease and a previously treated infection, which can be critical for determining whether treatment is required.”

In parallel, the system is also being developed to detect viral haemorrhagic fevers using molecular amplification techniques. Rather than relying on conventional PCR testing, which requires repeated temperature cycling to replicate genetic material, the project uses isothermal amplification, allowing the reaction to take place at a constant temperature.

For point-of-care diagnostics, that distinction is important. PCR systems typically require precise thermal cycling equipment, which adds complexity and limits where tests can be performed.

“We perform the assay at a single temperature using isothermal amplification,” Spasic explains. “Because the reaction runs under constant conditions, it removes the need for the complex temperature cycling used in PCR systems, which makes the technology much easier to integrate into a portable diagnostic device.”

Once the reaction has taken place, the result can be translated into a simple visual signal within the device itself. The system is designed to display the outcome in a familiar format, similar to a pregnancy test. “It’s a qualitative result,” Lammertyn adds. “So the answer is yes or no, simple to understand.”

Making diagnostics accessible
While the technology behind FORTIFIEDx is complex, the user experience is designed to be straightforward. The device is applied to the skin like a patch. After sampling, the internal processes run automatically without further intervention and the result becomes visible after a short period. The aim is to reduce the level of training required to perform diagnostic tests as even relatively simple rapid tests can prove challenging for untrained users, as Van den Bossche notes.

“If you think about COVID tests, you had to add drops of buffer,” she says. “Did I put one drop too many? Did I do it correctly? It creates questions.”

Handling blood samples adds another layer of difficulty. “It seems easy, but when you have to do it with lay users, it becomes much more complex,” she explains.

By automating the entire process, including sample metering and reagent mixing, the FORTIFIEDx system could make testing more robust and reliable. “It would create more accurate test results compared to current rapid diagnostic tests when they are used by non-experts,” she says.

Real-world deployment
Taken together, these design choices are intended to make the device simple to use outside traditional laboratory settings. By integrating sampling, fluid handling and analysis into a single patch, the system is designed to minimise the need for specialised training or complex equipment – an important consideration for point-of-care testing, particularly in underserved or remote regions.

The project is currently just over halfway through its development phase. According to Lammertyn, the individual components of the system are already taking shape. “We have developed many of the different modules,” he says. “Now the challenge is to bring them together and test them in real-life situations.”

Those tests will take place not only in European laboratories but also in settings where access to diagnostics is limited. “The validation will also be done in developing countries, which will be supported by our ITM partner (Belgium) and CNFRSR partner from Guinea” he says.

Even with a successful prototype, however, significant work will remain before the technology reaches patients. Medical devices must undergo extensive regulatory approval processes, particularly when they incorporate invasive components such as microneedles. “We have to prove safety, reliability, sensitivity and specificity,” Lammertyn explains. “Because we integrate microneedles, the regulatory pathway becomes more complex.”

Navigating these regulatory requirements is a complex process that extends beyond the laboratory and this project scope. Universities typically work with specialist regulatory consultants and industrial partners who can help manage the approval pathway and prepare emerging technologies such as FORTIFIEDx for eventual market deployment. Despite these challenges, however, the project team is optimistic about its long-term potential. “Our aim is to deliver a strong proof of concept,” Spasic says. “A prototype that clearly demonstrates the potential of this approach.”

If successful, FORTIFIEDx could represent an important step towards a new generation of decentralised diagnostics. By bringing sampling, analysis and result delivery together in a single device, the technology is designed to make reliable testing possible in settings where laboratory infrastructure, specialist equipment or trained personnel may not be readily available.

That could have significant implications for how infectious diseases are detected and managed, particularly in communities where access to healthcare services remains limited and where diagnosis needs to happen closer to the patient rather than in distant laboratories. “In many settings, people have to travel long distances to reach a healthcare facility,” Lammertyn says. “If they do that and they don’t receive the result immediately, they may never come back.”

Providing immediate results at the point of care could change that dynamic entirely, enabling earlier diagnosis, faster treatment decisions and reducing the risk of transmission. “And that,” he adds, “is where this technology can bring real value.”