COPRO-MOLECULAR CHARACTERIZATION OF CRYPTOSPORIDIUM SPP. AND GENOTYPES AMONG EGYPTIAN CHILDREN
Insights
Cryptosporidium infection in children was detected using ELISA and nested PCR (nPCR). Nested PCR identified more cases, revealing two genotypes, with genotype 1 being more prevalent, suggesting a higher risk from human sources.
Area of Science:
- Medical Parasitology
- Molecular Diagnostics
- Public Health
Background:
- Cryptosporidium is a significant cause of childhood diarrhea globally.
- Accurate detection and genotyping are crucial for understanding transmission dynamics and implementing control measures.
Purpose of the Study:
- To compare the diagnostic performance of ELISA and nested PCR (nPCR) for Cryptosporidium detection in children.
- To identify Cryptosporidium genotypes circulating in symptomatic and asymptomatic children in Cairo.
- To investigate the potential sources of infection based on genotype prevalence.
Main Methods:
- Stool samples from 182 diarrhoeic and 100 asymptomatic children were analyzed using ELISA and nPCR targeting the COWP gene.
- Positive nPCR products were genotyped using RsaI restriction fragment length polymorphism (RFLP).
Main Results:
- Nested PCR showed higher detection rates (22.3%) compared to ELISA (11.3%).
- ELISA demonstrated 47.6% sensitivity and 99.1% specificity against nPCR.
- Two genotypes were identified: genotype 1 (88.3%) and genotype 2 (11.7%), with genotype 1 being predominant.
Conclusions:
- Nested PCR is a more sensitive method for Cryptosporidium detection in children than ELISA.
- The high prevalence of genotype 1 suggests a greater contribution of human-to-human transmission in this population.
- Further research into the epidemiology and transmission of Cryptosporidium genotypes in children is warranted.
Abstract:
Stool samples from 182 diarrhoeic (symptomatic) children and 100 apparently healthy (asymptomatic) children, matched for age, from Aboul-Reesh Cairo University Pediatrics Hospital were examined by ELISA and by nPCR (targeting COWP gene) for the detection of Cryptosporidium. The demographic and environmental data of the diarrhoeic group was recorded. The PCR amplified product of positive cases was then subjected to RFLP by digesting it with the restriction enzyme RsaI. The obtained fragments were resolved by electrophoresis and the bands were visualized and characterized versus a standard. ELISA results demonstrated a prevalence rate of 13.2% (24/182) among diarrhoeic group, and 8% (8/100) among non-diarrheic group, with overall detection rate of 11.3% (32/282). Higher rates of detection were obtained by nested PCR assay among diarrhoeic group 25.8% (47/182) and 16% (16/100) among non-diarrhoeic group with overall detection rate of 22.3% (63/282). Considering nPCR as the reference method, ELISA had a sensitivity of 47.6% and a specificity of 99.1%. RsaI digestion of nPCR product of COWP gene revealed the presence of 2 genotypes: genotype 1 with 4 bands (34, 106, 125 and 285) and genotype 2 in which 3 bands (34, 106 and 401). Among the 63 cases with cryptosporidiosis, 53 (88.3%) had genotype 1, and 7 (11.7%) had genotype 2. The higher prevalence of genotype 1 suggests a relatively greater risk of human source of infection than zoonosis.
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