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Communication Dans Un Congrès Année : 2021

Development of an aptamer based test for Trichinella detection

Résumé

Aim: Trichinellosis is a zoonotic illness transmitted through the consumption of raw or undercooked meat products infected with the parasitic nematode Trichinella spp. Excluding its brief existence as a free-living migratory larva, Trichinella spp. resides primarily within the confines of its host’s muscle cells. In this study, we employ an innovative and novel larval-based selection method to produce a set of Trichinella spiralis specific aptamers for use in an accurate diagnostic method for animal and human infection. Methods: A magnetic stirrer method of artificial digestion and microscopy were used for the recovery and quantification of Trichinella spiralis muscle larvae (ML). Aptamers were selected from a synthetic single-stranded DNA (ssDNA) library composed of 1013-1016 random sequences by an iterative in vitro selection process termed Systematic Evolution of Ligands by EXponential enrichment (SELEX). Additionally, the selection process and sequence pool quality were monitored by gel electrophoresis. Results: Currently, T. spiralis ML have been successfully isolated from experimentally infected mice and preserved in 70 % ethanol. Furthermore, 5 rounds of SELEX have been successfully performed based on gel electrophoresis results, indicating the presence of high quality aptamer sequences. Conclusions: In light of these results, additional cycles of selection are to be performed while applying new methods to obtain high quality aptamer sequences. Furthermore, melting curve analysis and confocal microscopy will be implemented to more accurately evaluate aptamer pool diversity and specificity. Finally, T. spiralis specific aptamers could act as promising candidates as molecular recognition elements in diagnostic assays or for therapeutic purposes.
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Dates et versions

anses-04140796 , version 1 (26-06-2023)

Identifiants

  • HAL Id : anses-04140796 , version 1

Citer

Noah Brosseau, Isabelle Vallée, Anne Mayer-Scholl, Momar Ndao, Grégory Karadjian. Development of an aptamer based test for Trichinella detection. 3rd Annual Scientific Meeting of the One Health EJP, Jun 2021, Copenhagen, Denmark. ⟨anses-04140796⟩
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