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. 2011 Winter;4(1):29-33.

Genotyping of Cryptosporidium spp. in clinical samples: PCR-RFLP analysis of the TRAP-C2 gene

Ehsan Nazemalhosseini Mojarad  1 , Akbar Keshavarz  1 , Niloofar Taghipour  1 , Ali Haghighi  1 , Bahram Kazemi  1 , Amid Athari  1
Affiliations

Genotyping of Cryptosporidium spp. in clinical samples: PCR-RFLP analysis of the TRAP-C2 gene

Ehsan Nazemalhosseini Mojarad et al. Gastroenterol Hepatol Bed Bench. 2011 Winter.

Abstract

Aim: The aim of the present study was to determine the species and genotypes of Cryptosporidium spp. among children with diarrhea by PCR-RFLP using the TRAP-C2 gene.

Background: Cryptosporidium is a globally distributed protozoan parasite and one of the most common causes of infection and diarrhea in humans.

Patients and methods: Four hundred and sixty nine stool samples were collected from children less than 12 years with diarrhea who had been referred to Pediatrics Medical Centers in Gazvin provinces. The presence of Cryptosporidium oocysts was determined by Ziehl-Neelsen acid fast staining, then, genomic DNA was extracted from positive samples and nested PCR-RFLP was performed to amplify the TRAP-C2 gene.

Results: The overall prevalence of Cryptosporidium infection in children was 2.5 %. Results of nested PCR amplification showed that of 12 positive children samples, 10 (83.3%) were belonged to C. parvum, followed by C. hominis in 1 (8.3%) and mixed infection in 1 isolate (8.3%).

Conclusion: This study showed that Cryptosporidium parvum (the zoonotic genotypes) is more prevalent than other Cryptosporidium species in children from this area. This suggests that zoonotic transmission is the main mode of transmission of Cryptosporidium infection in Iran.

Keywords: Cryptosporidium; Genotypes; TRAP-C2 gene.

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Figures

Figure 1
Figure 1
Molecular diagnosis of Cryptosporidium spp. by a nested PCR based on TRAP-C2 gene. Lane 1: 100 bp
Figure 2
Figure 2
Gel electrophoresis of Cryptosporidium species with TRAP-C2 gene based on PCR-RFLP technique by digestion of the secondary PCR products in clinical isolates with HaeIII, BstEII enzyme. Lane 1, 4, 5, 6, 8: C. parvum, Lane 2, 3: C. huminis, Lane 7: mixed infection both genotype
See this image and copyright information in PMC

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