RESUMO
Microsporidia and Cryptosporidium species are prominent agents of enteritis, capable of causing severe chronic diarrhoea in children, immunocompetent and immunocompromised individuals around the world. It is not possible to identify the parasites at species level solely on the basis of microscopy. The aim of the present study was to identify and characterize the species of Microsporidia and Cryptosporidium in immunocompetent humans with GI disturbances by nested PCR-RFLP, sequencing, and phylogenetic analysis. Fresh frozen and fresh paraffin-embedded biopsy specimens of the duodenum, jejunum, ileum and the cecum of 110 patients were examined. Genomic DNA was extracted from all bowel biopsies. Nested PCR targeting the 18S rRNA gene was performed by restriction endonuclease digestion of the PCR product followed by nucleotide sequencing and phylogenetic analysis. A total of three patients with chronic diarrhoea were positive for Microsporidia and Cryptosporidium spp. Species analysis showed the presence of C. parvum and E. bieneusi in two and one samples, respectively. This is the first PCR confirmation of the presence of E. bieneusi and C. parvum in a bowel biopsy of immunocompetent individuals in Iran. This study revealed that PCR, sequencing, and phylogenetic analysis are very powerful tools for the precise species identification of these pathogens.
Assuntos
Criptosporidiose/parasitologia , Cryptosporidium/genética , Intestinos/patologia , Intestinos/parasitologia , Microsporídios/genética , Microsporidiose/parasitologia , Filogenia , Biópsia , Criptosporidiose/epidemiologia , Cryptosporidium/classificação , Humanos , Irã (Geográfico)/epidemiologia , Microsporídios/classificação , Microsporidiose/epidemiologia , Reação em Cadeia da Polimerase/métodos , RNA Fúngico/genética , RNA de Protozoário/genética , RNA Ribossômico 18S/genéticaRESUMO
We report a case visceral leishmaniasis with disseminated cutaneous leishmaniasis caused by Leishmania tropica in southern Iran. Typing of this parasite was performed by a species-specific polymerase chain reaction and isoenzyme electrophoresis.
Assuntos
Leishmania tropica/isolamento & purificação , Leishmaniose Cutânea/parasitologia , Leishmaniose Visceral/parasitologia , Adolescente , Anfotericina B/uso terapêutico , Animais , Antiprotozoários/uso terapêutico , Feminino , Humanos , Interferon gama/uso terapêutico , Irã (Geográfico) , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Visceral/tratamento farmacológico , Meglumina/uso terapêutico , Antimoniato de Meglumina , Compostos Organometálicos/uso terapêutico , Fosforilcolina/análogos & derivados , Fosforilcolina/uso terapêuticoRESUMO
BACKGROUND: The causative agent of visceral leishmaniasis (VL) in Iran is Leishmania infantum (L. infantum) (Mediterranean type) and its major reservoir host is the dog. OBJECTIVE: To compare the serological methods including direct agglutination test (DAT), indirect immunofluorescent-antibody test (IFA) and enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of endemic strain of L. infantum. METHODS: 61 blood samples from VL patients referred to Shiraz hospitals and 49 blood samples from control group were collected. Native strain of the parasite isolated from a VL patient from the region was cultured and characterized. Antigens from this L. infantum parasite were used in ELISA and IFA system. RESULTS: Anti-Leishmania antibody was detected in 43 (70.5%), 49 (80.3%) and 51(83.6%) cases using DAT, IFA and ELISA, respectively. Based on these results, sensitivity and specificity of DAT was found to be 70.5% and 100%, respectively. Sensitivities of IFA and ELISA in diagnosis of VL were 80.3% and 83.6% and their specificity was 90.5%. CONCLUSION: Results of this study showed that DAT and ELISA have the highest specificity and sensitivity in diagnosis of VL. DAT is a simple, cost-effective and field applicable test. Thus, it can be recommended for early and accurate diagnosis of VL, especially in regions where malaria, brucellosis and tuberculosis are prevalent.
