title: Innovative molecular diagnosis of Trichinella species based on β-carbonic anhydrase genomic sequence.
abstract: Trichinellosis is a helminthic infection where different species of Trichinella nematodes are the causative agents. Several molecular assays have been designed to aid diagnostics of trichinellosis. These assays are mostly complex and expensive. The genomes of Trichinella species contain certain parasite-specific genes, which can be detected by polymerase chain reaction (PCR) methods. We selected β-carbonic anhydrase (β-CA) gene as a target, because it is present in many parasites genomes but absent in vertebrates. We developed a novel β-CA gene-based method for detection of Trichinella larvae in biological samples. We first identified a β-CA protein sequence from Trichinella spiralis by bioinformatic tools using β-CAs from Caenorhabditis elegans and Drosophila melanogaster. Thereafter, 16 sets of designed primers were tested to detect β-CA genomic sequences from three species of Trichinella, including T. spiralis, Trichinella pseudospiralis and Trichinella nativa. Among all 16 sets of designed primers, the primer set No. 2 efficiently amplified β-CA genomic sequences from T. spiralis, T. pseudospiralis and T. nativa without any false-positive amplicons from other parasite samples including Toxoplasma gondii, Toxocara cati and Parascaris equorum. This robust and straightforward method could be useful for meat inspection in slaughterhouses, quality control by food authorities and medical laboratories.
authors.affiliation: School of Medicine, University of Tampere, FI-33520, Tampere, Finland.|BioMediTech, University of Tampere, FI-33520, Tampere, Finland.|Fimlab Laboratories Ltd and Tampere University Hospital, Biokatu 4, FI-33520, Tampere, Finland.
authors.forename: Reza
authors.initials: R
authors.lastname: Zolfaghari Emameh
authors.affiliation: School of Medicine, University of Tampere, FI-33520, Tampere, Finland.
authors.forename: Marianne
authors.initials: M
authors.lastname: Kuuslahti
authors.affiliation: Department of Veterinary Biosciences, Faculty of Veterinary Medicine, University of Helsinki, FI-00014, Helsinki, Finland.
authors.forename: Anu
authors.initials: A
authors.lastname: Näreaho
authors.affiliation: Department of Veterinary Biosciences, Faculty of Veterinary Medicine, University of Helsinki, FI-00014, Helsinki, Finland.
authors.forename: Antti
authors.initials: A
authors.lastname: Sukura
authors.affiliation: School of Medicine, University of Tampere, FI-33520, Tampere, Finland.|Fimlab Laboratories Ltd and Tampere University Hospital, Biokatu 4, FI-33520, Tampere, Finland.
authors.forename: Seppo
authors.initials: S
authors.lastname: Parkkila
mesh_terms: D000818:Animals; D002256:Carbonic Anhydrases / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D008460:Meat / Q000469:parasitology*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D017160:Trichinella spiralis / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D014235:Trichinellosis / Q000175:diagnosis*
chemical_list: D017931:DNA Primers; D002256:Carbonic Anhydrases