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Lab-on-a-chip microfluidic device based on plasmonic driven thermal nanobiosensing for rapid detection of Salmonella in chicken meat.

Periodic Reporting for period 2 - HEATSENS_S (Lab-on-a-chip microfluidic device based on plasmonic driven thermal nanobiosensing for rapid detection of Salmonella in chicken meat.)

Reporting period: 2017-06-01 to 2019-02-28

"HEATSENS® is an innovative and disruptive nanobiosensing technology owned by NANOIMMUNOTECH, that allows designing ultra-sensitive and easy-to use devices for a rapid detection of analytes of interest. This revolutionary biosensing platform is based on the heat generated by gold nanoprisms when they are irradiated with an external light source. At the end of the project HEATSENS®_S, NANOIMMUNOTECH has achieved the fully integration of this technology in a device to detect Salmonella in chicken meat within one working day.
Salmonella is a bacterium that can cause an illness called salmonellosis in humans. Salmonellosis is the second most common foodborne disease after campylobacteriosis in the EU and Salmonella is an important cause of foodborne outbreaks. In 2017, Member States reported 91,662 cases in humans, and according to the words of Marta Hugas, Chief Scientist at European Food Safety Authority (EFSA), after years of significant progress in reducing the burden of foodborne illnesses in the EU, especially Salmonella, the situation has now stalled, and increased efforts are needed to push the figures further down, ( EFSA (European Food Safety Authority) https://www.efsa.europa.eu/en/press/news/181212) . On the other hand, recently EFSA reported that there is conclusive evidence that an increased stocking density, larger farms and stress result in increased occurrence, persistence and spread of Salmonella in laying hen flocks, (EFSA Journal (amended 8 April 2019) https://efsa.onlinelibrary.wiley.com/doi/epdf/10.2903/j.efsa.2019.5596).
To protect consumers from Salmonella, the EU has adopted a regulation that agri-food companies must comply. European Regulation is the most restrictive, being the threshold for these controls extremely low: meat is considered to be contaminated if there is one colony-forming unit (CFU) in 25 mg of meat, and the total absence of bacteria must be guaranteed for the rest of samples, tools and surfaces in contact with the animals, under the ISO 6579-1:2017 standard: Microbiology of the food chain — Horizontal method for the detection, enumeration and serotyping of Salmonella — Part 1: Detection of Salmonella spp, (ISO (the International Organization for Standardization) https://www.iso.org/obp/ui/#iso:std:iso:6579:-1:ed-1:v1:en) .
Therefore, response time to get the result is very relevant for the producers (end-users and costumers of HEATSENS®): results should arrive on time to treat the animals, and mainly they should arrive before the product’s release. This need has been solved by HEATSENS®_S, being the only method to detect salmonella in chicken meat within the same working day.
As already mentioned, NANOIMMUNOTECH worked to apply HEATSENS® technology in the agri-food sector, developing the final HEATSENS®_S Product, a biosensor based on HEATSENS® technology which solves a need in one specific market with great growth potential: the detection of Salmonella in chicken meat in a faster way than current market solutions, giving results within the same working day.
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From the beginning of the project NANOIMMUNOTECH has carried out the scale-up and optimization of the nanoparticles´ production. Gold nanoprisms synthesis besides to their functionalization with quality control were optimized and scaled-up from a lab-scale to a large-scale production ensuring the amount needed for the supply and performance of the tests in the different devices. Another key task, the chip layout besides to its chemical functionalization have been also optimized allowing its scale-up. The functionalization protocol of the microfluidic chip was established for the linear chip layout. All the relevant parameters of the immunoassay involved in the performance of the microfluidic chip were optimized.
During the final period the tasks 1.4 1.5 and 1.6 included in the Work Package 1 (first prototypes´ production) have been successfully accomplished. Firstly, MICROLIQUID as manufacturer subcontracted has provided the manufacturing strategy to assemble completely the prototypes and the fabrication protocols for it, and a first beta production of four instruments was accomplished, after a global validation and several quality control processes of the different core components of the prototype. This validation allowed the obtaining of HEATSENS®_S final prototype according the compliance of all specifications required including those necessary for CE certification process. Finally, NANOIMMUNOTECH carried out the demonstration to its customers, allowing to obtain significant results and thus, to validate the technology in a global way by comparison to certified methodology (conventional immunoassays, microbiological culture and molecular biology techniques). Regarding dissemination and communication actions, NANOIMMUNOTECH fulfilled the objectives set out by their Marketing Plan for starting the next commercialisation stage once HEATSENS® is ready to market.
HEATSENS_S Product is composed of consumables (chips to analyse Salmonella – Test_S) and a device that reads them (HEATSENS® device). Companies will acquire HEATSENS® device (probably under financial leases) and will buy as many consumables as they need for their tests (Test_S).
Thanks to ultrasensitive HEATSENS® technology, final users will get the results of Salmonella within one working day, for an affordable cost per test. That’s is because HEATSENS®_S can detect Salmonella at a much lower concentration in the sample, reducing the pre-treatment stage time. The time of analysis of Salmonella will be 90 min after loading the Salmonella detection test (Test_S) on the HEATSENS® device.
Retailers demand producers to use highly sensitive methods to detect Salmonella, because it will bring benefits to final chicken consumers, who are anyone of us, our children or family. The more sensitive the method is, the lower the risk of being infected by Salmonella. This issue is crucial for retail stores, which are the point of sale, namely the intermediary between integrated companies and the final customer (for example, supermarkets). For them, sensitivity means money: sensitivity allows quick detection, and that means having the product more time saleable in the outlet.
NANOIMMUNOTECH is the sole exploiter of HEATSENS® technology. HEATSENS® development and commercialisation is the core business of the Company. HEATSENS®_S exploitation will be itself profitable for the Company. However, it is only the beginning.
Once that the HEATSENS® device is ready to market, it will enable the detection of different analytes by changing the consumables. Furthermore, the development of different consumables (Tests) will be carried out within a short period of time, estimated between 6-12 months, mainly depending on the type of analyte (microorganisms, proteins, oligonucleotides, etc.) and the matrix (meat, chocolate, serum, blood, urine, faeces or water) in which it is found.
The product is expected to be launched in 2019 and the Sales and Marketing team are working on identifying the most adequate strategy of launch. At the same time, the financial team is working on the final strategy according the expected forecasts of the Company numbers.