Innovative molecular diagnosis of T richinella species based on β‐carbonic anhydrase genomic sequence

Trichinellosis is a helminthic infection where different species of T richinella 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 T richinella 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 T richinella larvae in biological samples. We first identified a β‐CA protein sequence from T richinella spiralis by bioinformatic tools using β‐CAs from C aenorhabditis elegans and D rosophila melanogaster. Thereafter, 16 sets of designed primers were tested to detect β‐CA genomic sequences from three species of T richinella, including T . spiralis, T richinella pseudospiralis and T richinella 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 T oxoplasma gondii, T oxocara cati and P arascaris equorum. This robust and straightforward method could be useful for meat inspection in slaughterhouses, quality control by food authorities and medical laboratories.