ABSTRACT
To determine the sensitivity and specificity of routine screening methods for cryptosporidiosis, three methods including conventional modified Ziehl-Neelsen (MZN), direct fluorescent-antibody (DFA) and Nested-PCR assay compared together. To this end, their ability to identify the low concentrations of Cryptosporidium spp. oocysts in children fecal samples was evaluated. The sample population of this study was children under 12 years old who had diarrhea and referred to pediatric hospitals in Tehran, Iran. 2,510 stool specimens from patients with diarrhea were screened for Cryptosporidium oocysts by concentration method and MZN. To determine sensitivity and specificity, Nested-PCR and DFA were performed on 30 positive and 114 negative samples which previously had been proved by MZN. By using the microscopic method, DFA assay and PCR analysis, a total of 30 (1.2 %), 28 (1.1 %) and 32 (1.27 %) positive samples were detected respectively. According to the results, the sensitivity, specificity, and positive and negative predictive values of the Nested-PCR assay were 100 %, compared to 94, 100, 100, and 98 %, respectively, for MZN and 87.5, 100, 100, and 96 %, respectively, for DFA. Results of the present study showed that the Nested-PCR assay was more sensitive than the other two methods and laboratories can use the Nested-PCR method for precise diagnosis of Cryptosporidium spp. However, regarding the costs of Nested-PCR and its unavailability in all laboratories and hospitals, MZN staining on smears has also enough accuracy for Cryptosporidium diagnosis.
ABSTRACT
A comprehensive survey assessing the presence of Acanthamoeba was conducted on 50 samples from water sources in parks and public squares from 22 municipal districts of Tehran, Iran. The prevalence and genotypes of Acanthamoeba were determined by PCR and the PCR fragments of ribosomal RNA genes sequenced. Sixteen (32%) samples were positive for Acanthamoeba spp. Sequence analysis revealed that the positive isolates belonged to the T4 and T5 genotypes. Fourteen isolates (87.5%) were T4, and two (12.5%) were T5. Acanthamoeba may be a problematic organism for contact lens wearers and for immunocompromised individuals. In Iran, Acanthamoeba keratitis has increased in recent years, mainly due to poor hygiene in contact lens wearers. A thorough survey for the prevalence of this amoeba could have a significant role in prevention of disease. This is the first report of the T5 genotype from water in recreational areas of Tehran.
