Molecular identification and characterization of Cryptosporidium spp. among pediatric and food-handlers subjects in Qatar

Abstract

World Health Organization (WHO) has identified Cryptosporidium spp. as globally the most common diarrhea-causing protozoan. Its transmission occurs mainly through ingestion of water or food contaminated with the feces of an infected human or animal. Infected food handlers are also a major Cryptosporidium contamination risk. A wide diversity of Cryptosporidium spp. and subtypes infect humans and due to the host specificity of each species, the use of molecular techniques and genotyping of subject’s stool samples will help in identifying contamination source. Molecular prevalence was investigated using real-time PCR targeting a 214 base pairs fragment of 18s-rRNA. The analyzed populations consist of hospitalized diarrheic pediatrics and asymptomatic food handlers. The positive samples by RT-PCR were subjected to species identification by PCR-RFLP and GP60 gene sequencing. The parasite was detected by RT-PCR in 15% of the children and 4% of the food-handlers. Genotyping of Cryptosporidium spp. revealed a predominance of Cryptosporidium parvum. Sequence analysis of the GP60 locus in the present study has identified only C. parvum subtype family IId with mostly the IIdA20G1 subtype among the two populations. This study provides supplementary information for implementing prevention and control strategies to reduce the burden of these pediatric protozoan infections.