2155 Avenue Guy, Montreal, Québec, Canada

Max-Dohrn-Straße, Berlin, Berlin, Germany

14 Rue Pierre et Marie Curie, Paris 5e Arrondissement, Île-de-France, France

OHEJP APTATRICH project logo

The Project #APTATRICH

Start: November 2019
Duration: 3 years
Domain: Foodborne Zoonoses
Members: ANSES- France, BfR- Germany
Contact: Dr Gregory Karadjian

AptaTrich: Development of an aptamer-based test for Trichinella detection

Trichinellosis is a zoonosis caused by the consumption of raw or undercooked meat of animals (mainly pigs, wild boars, horses) infected with the nematode Trichinella spp. Over the past decades, human trichinellosis incidence has decreased in the EU due to improvement of pig farming conditions and to stringent control measures implemented by controls at the slaughterhouse. The cost of trichinellosis control in pigs is estimated at 220 millions euros per year in EU. To date, Trichinella spp. remain in the top three of prioritized foodborne parasites in Europe (Bouwknegt et al., 2018). This parasite is still of major public health and economic importance at international level (Codex Alimentarius).

EFSA identified the type of production system as a main risk factor for Trichinella infections, and that the risk of Trichinella infection in pigs from officially recognised controlled housing conditions (ORCHC) is negligible. Therefore, EU regulation 2015/1375 introduced possible derogations from the necessity of testing each slaughter pig, if specific conditions are met. In order to verify that Trichinella is truly absent in such a population and to identify changes in disease prevalence at an early stage, monitoring programmes are recommended (EU 2015/1375). To date, serological methods have been identified, but are fraught with problems concerning specificity (false positives) and detection of infection at an early stage.

Due to the very low Trichinella prevalence in pigs from ORCHC, a test specificity bordering 100% is needed, as false positive samples would need to be retested with a second serological method (e.g. Western Blot). Such tests can only be performed by specialised laboratories, making the testing logistics more complicated and expensive. Therefore, new diagnostic methods with higher specificity and earlier detection are needed for prevention at the slaughterhouse and to improve human disease detection.

When a human outbreak occurs, the importance of early onset therapy is clearly needed as this avoids complications and reduces the patient’s parasitic load. To date, human diagnosis is based on the detection of specific IgG that appear as early as 10-20 days post infection. However, antibody titers are highly variable depending on the individuals, the initial parasite load and the infecting species. Numerous tests have been described but the most efficient systems remain the ELISA based on the use of excretion / secretion antigens (Ag E/S) and Western blot performed with this antigen. The Western blot makes it possible to obtain the best specificity; however, some cross-reactions are observed with anisakiases and bilharziases. The ELISA can be positive as early as the second week post infection, but generally, the serological reactions are clearly positive only one month post infection. This is often too late for anthelminthics to effectively reach the nematodes, as tissue cyst envelop the worm in less than two weeks. Therefore, new diagnostic methods with higher specificity and earlier detection times in meats are needed for prevention and in human serum for detection of disease.

Aptamers are synthetic nucleic acids that fold into unique three-dimensional conformations capable of binding a target with remarkable affinity and specificity. These structures specifically bind to pathogen antigens, thus combining the ease of serological sampling, and the direct detection of the presence of the pathogen. Aptamers have successfully been used for the detection of parasites in fresh produce, including in aptamer-based biosensors. The development of an aptamer based detection system for Trichinella would circumvent the caveats associated with serological testing and enable specific and early detection in both human diagnostics and Trichinella monitoring programmes in pigs. Further, the technique can be combined with aptamers directed against other pig diseases (such as Toxoplasma, Salmonella etc), making a much wider future application possible.

Project Aims

In this study, we employ an innovative and novel larval-based selection method entitled whole-larva Systematic Evolution of Ligands by Exponential enrichment (SELEX) to produce a set of Trichinella spiralis specific aptamers for potential therapeutic and diagnostic applications. In addition, biomarkers of infection are to be identified, evaluated and targeted by protein SELEX to develop a novel aptamer-based tool for accurate and sensitive detection in humans and animals.


Whole larva SELEX was used to select aptamer sequences capable of binding the muscle larva (ML) stage of Trichinella spiralis. Sequence enrichment was qualitatively monitored using qPCR-based melting curve analysis by assessing the gradual reduction in sequence pool diversity throughout the SELEX process. Furthermore, Next-generation sequencing (NGS) and bioinformatics analysis were used to quantitatively monitor sequence evolution with a higher sensitivity.

 Results obtained

  • A whole-larva SELEX method was meticulously developed and optimized
  • Next-generation sequencing (NGS) and bioinformatics analysis has confirmed the selection of a 11-mer structural motif that is shared among several sequence families in three independent SELEX experiments.
  • Confocal fluorescence microscopy has confirmed the ability of most selected aptamers to bind the surface of T. spiralis ML. Some sequences appear to bind more consistently than others.

Project Assets

Poster Presentation at Journee des Sciences de la Vie et de la Sante a Creteil, Salons de l’Aveyron, Paris, France. 19th Oct 2022.

Oral Presentation at Journée de la recherche ENVA, Ecole Vétérinaire d’Alfort, Ile-de-France, France. 20th September, 2022.

Poster Presentation at Non-coding Genome 11th Edition 2022, Institut Curie, Paris, France. 11-18th May, 2022.

Oral Presentation – 3 minute thesis presentation at OHEJP ASM 2022, Orvieto, Italy. 11-13th April 2022.

Oral Presentation at Journee des Sciences de la Vie et de la Sante a Creteil, Creteil, Ile-de-France, France. 16th February 2022.

Brosseau, N. E., Vallée, I., Mayer-Scholl, A., Ndao, M., Karadjian, G. (2023). Aptamer-Based Technologies for Parasite Detection. Sensors.  23(2), 562. DOI: https://doi.org/10.3390/s23020562

Le Dortz, L.L., Rouxel, C., Leroy, Q., Brosseau, N., Boulouis, H-J, Haddad, N., Lagrée, A-C., Deshuillers, P.L. (2022). Optimized Lambda Exonuclease Digestion or Purification Using Streptavidin-Coated Beads: Which One Is Best for Successful DNA Aptamer Selection? Methods and Protocols. 5(6), 89. DOI: https://doi.org/10.3390/mps5060089

Deliverable 1 : ‘T. spiralis muscle larvae are fixed in ethanol’

Final PhD Thesis Report

OHEJP APTATRICH project noah brosseau phd student photo

Noah Brosseau

About me:
I am a 25 year old Montreal native currently living with two of my closest friends in Verdun, Quebec. After graduating from McGill University’s Life Sciences program, I decided to take some time off from school to explore my interests. After working for many months as a valet driver at a hotel in Montreal’s old city, my profound interest in neglected tropical diseases (NTD’s) lead me to the Research Institute of McGill University Health Centre (RI-MUHC). During my time I worked on a project entitled “Detection of Fusobacterium nucleatum subspecies in saliva and colorectal cancer tissue using matrix-assisted laser desorption ionisation mass spectrometry (MALDI-MS)”. Outside of school I very much enjoy cooking, photography, and making music with friends.

What motivated me to do a PhD:
My motivations for the completion of this thesis are to have a long lasting and influential impact on both the lives of the people that are most at risk to <em>Trichinella</em> infection and the scientific community. My future professional aspirations in the field of global and public health coincide with the goals and methods of this thesis. Eventually I aim to focus my work on the Neglected tropical diseases (NTDs) that are so heavily burdening the third world countries of the Global South. I believe that this diverse group of tropical infections, most common in low-income populations, is partly to blame for the inability of these countries to develop and prosper. Aiding in the development of a method for the early detection of <em>Trichinella spiralis</em> could ultimately lead to similar aptamer-based technologies for the detection of other serious illnesses.



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