Molecular differentiation of cattle Sarcocystis spp. by multiplex PCR targeting 18S and COI genes following identification of Sarcocystis hominis in human stool samples

• Published in: Food and waterborne parasitology 2020-03, Vol.18, p.e00074-e00074, Article e00074
• Main Authors: Rubiola, S., Civera, T., Ferroglio, E., Zanet, S., Zaccaria, T., Brossa, S., Cipriani, R., Chiesa, F.
• Format: Article
• Language: English
• Subjects: Cattle; COI (COX1) gene; cytochrome-c oxidase; definitive hosts; feces; genes; humans; intestines; meat; mitochondria; Multiplex PCR; nucleotide sequences; parasites; patients; polymerase chain reaction; Protozoa; ribosomal RNA; risk; Sarcocystis; Sarcocystis hominis; Sarcocystis spp; sarcocystosis; Zoonosis
• ISSN: 2405-6766; 2405-6766
• DOI: 10.1016/j.fawpar.2020.e00074

Sarcocystis spp. are protozoan parasites which can infect a wide range of vertebrates, including humans; the latter can act as definitive hosts for two cattle Sarcocystis spp.: Sarcocystis hominis and Sarcocystis heydorni. Reports of intestinal sarcocystosis are well documented in the literature, but PCR-based methods have been scarcely used to identify Sarcocystis species in human stools, and have been limited to the molecular analysis of 18S ribosomal RNA (18S rRNA) gene sequences. Since the mitochondrial cytochrome c oxidase subunit I (COI) gene is one of the most promising tools for distinguishing between closely related Sarcocystis spp., and taking into account the lack of publicly available S. hominis COI sequences, in the present study we obtained the first partial COI sequence of S. hominis from human stool samples of patient with gastrointestinal symptoms. We designed specific COI primers to develop a multiplex PCR method for the identification of Sarcocystis spp. in cattle. The submission of the COI sequence described herein and the unambiguous identification of S. hominis through the application of the new multiplex PCR is important for determining the prevalence of this zoonotic Sarcocystis spp. in meat and the risk for consumers. •Human stool samples of patients with gastrointestinal symptoms were PCR-positive for Sarcocystis hominis.•The mitochondrial cytochrome c oxidase subunit I (COI) gene of S. hominis was amplified and sequenced.•A multiplex-PCR assay was developed to detect the presence of different cattle Sarcocystis in meat.•The unambiguous identification of S. hominis allows to estimate the prevalence in meat and the risk for the consumer.