Genotyping of Cryptosporidium-Positive Samples

Cryptosporidium species were determined using real-time polymerase chain reaction (PCR) analysis incorporating specific primers and probes for a C. parvum hypothetical protein and a C. hominis target for part of the GP60 gene [19]. These specific markers were chosen because C. parvum and C. hominis had been previously detected in the Netherlands during the 2012 increase in Cryptosporidium infections [17], and our study aimed to identify sporadic cases for cryptosporidiosis in the Netherlands. The duplex real-time PCR used is able to amplify a specific C. parvum sequence and a specific C. hominis sequence, allowing detection of coinfection with both species.