Assessment of the first commercial multiplex PCR kit (ParaGENIE crypto-micro real-time-PCR) for the detection of Cryptosporidium spp., Enterocytozoon bieneusi and Encephalitozoon intestinalis from fecal samples

Abstract Cryptosporidiumspp. Enterocytozoon bieneusiand Encephalitozoon intestinalisare opportunistic pathogens responsible for gastrointestinal diseases. We evaluated the ParaGENIE Crypto-Micro Real-Time PCR kit (Ademtech, France), the first CE-IVD compliant PCR assay available for these pathogens. This study was conducted blindly against a reference panel of 115 stool specimens including positive samples for Cryptosporidiumspp. ( n  = 48) and E. bieneusi ( n  = 38) as well as negative or positive samples for other parasites to test for cross-reactivity. An additional set of samples corresponding to 8 rare Cryptosporidiumspecies was also included. Discrepancies were evaluated with external in-house PCR tests. The ParaGENIE Crypto-Micro PCR assay displayed a sensitivity/specificity of 91.7%/100% and 97.3%/98.7% for Cryptosporidiumspp. and E. bieneusi , respectively, and was able to detect all 12 Cryptosporidiumspecies of the reference panel, including rare species. This new CE-IVD assay will facilitate the diagnosis of intestinal cryptosporidiosis and microsporidiosis, a major concern in immunocompromised patients and travelers.