Molecular characterization of Cryptosporidium isolates from humans by nested polymerase chain reaction – restriction fragment length polymorphism (nPCR-RFLP) analysis in Egypt

ABSTRACT

@#Cryptosporidium species is a group of protozoan parasites recognized as a cause of diarrhea with significant morbidity and mortality in industrialized and developing countries. Cryptosporidium hominis and Cryptosporidium parvum seem to be the most common causes of human cryptosporiodosis, however, other species are also found. Hence, the present study aimed to determine Cryptosporidium spp. infecting humans by nested PCR-RFLP targeting 18 sRNA gene in Minia Governorate, Egypt. A total of 300 diarrheic stool samples have been collected from inpatient and outpatient clinics of University Hospitals and Tropical Hospital, Minia Governorate, Egypt. One hundred twelve positive samples for Cryptosporidium infection (37.3%) were detected by nPCR while, 60 positive samples (20%) were detected by routine microscopy (p-value < 0.0004). The one hundred twelve positive samples detected by nested PCR were processed to RFLP. The RFLP yielded a typical restriction patterns for C. hominis in 73 (65.2%) cases, C. parvum in 25 (22.3%) cases, and C. meleagridis in 14 (12.5%) cases. C. hominis was more prevalent among cases of urban areas with negative history of animal contact in comparison to cases of rural areas (95.5% vs. 21.7%; p-value < 0.001). Moreover, there was a statistically significant association between C. hominis infection and drinking tap water. C. parvum was more prevalent among cases of rural areas than among those of urban areas (47.8% vs. 4.5%; p-value < 0.001) While C. meleagridis was only present among cases live in rural areas (30.4%; p-value < 0.001).