Assuntos
Transplante de Rim , Doenças Parasitárias/diagnóstico , Doenças Parasitárias/etiologia , Anti-Infecciosos/uso terapêutico , Cyclospora/classificação , Cyclospora/genética , Ciclosporíase/diagnóstico , Ciclosporíase/tratamento farmacológico , Ciclosporíase/etiologia , Enterocytozoon , Humanos , Masculino , Microsporidiose/diagnóstico , Microsporidiose/etiologia , Pessoa de Meia-Idade , Doenças Parasitárias/tratamento farmacológico , Viagem , Resultado do TratamentoRESUMO
BACKGROUND: Six species of the genus Entamoeba, i.e., E. histolytica, E. dispar, E. moshkovskii, E. polecki, E. coli, and E. hartmanii can be found in human stools. Among these, only E. histolytica is considered to be pathogenic, causing intestinal and extra-intestinal disease, but it is morphologically identical to E. dispar and E. moshkovskii. In general, E. polecki, E. coli, and E. hartmanii can be differentiated morphologically from E. histolytica, but some of their diagnostic morphologic features may overlap creating issues for the differential diagnosis. Moreover, the previous inability to differentiate among Entamoeba species has limited epidemiologic information on E histolytica. The objective of this study was to develop a rapid, high-throughput screening method using Luminex technique for the simultaneous detection and differentiation of Entamoeba species. METHODS: PCR amplification was performed with biotinylated Entamoeba sp 18S rRNA gene primers, designed to amplify a fragment ranging from 382 to 429 bp of the Entamoeba spp studied. Regions of this fragment that could differentiate among E. histolytica, E. moshkovskii, E. dispar, E. hartmanii and E. coli were selected to design hybridization probes to link to Luminex beads. The assay was standardized with cloned DNA samples of each species and evaluated with 24 DNA extracts from samples obtained from individuals diagnosed with these amebas in their stools. RESULTS: Using this approach we were able to correctly identify E. histoltyica, E. dispar, E hartmanni, E. coli and E. moshkovskii in all specimens studied. From twenty four samples tested by microscopy, PCR/DNA Sequencing and real-time PCR, 100% agreed with PCR-Luminex assay for identification of E. dispar, E. moshkovskii, E. hartmanni, E. histolytica, and E. coli. CONCLUSION: These results show that this method could be used in the diagnostic detection of Entamoeba spp in fecal samples. This diagnostic test was useful to clearly distinguish E histolytica from other species and also to strengthen epidemiologic data on Entamoeba spp.
Assuntos
Entamoeba/isolamento & purificação , Entamebíase/diagnóstico , Fezes/parasitologia , Programas de Rastreamento/métodos , Técnicas de Diagnóstico Molecular/métodos , Parasitologia/métodos , DNA de Protozoário/genética , Entamoeba/classificação , Entamoeba/genética , Entamebíase/parasitologia , Antropologia Forense , Humanos , Hibridização de Ácido Nucleico , Sondas de Oligonucleotídeos/genética , RNA Ribossômico 18S/genéticaRESUMO
The Phylum Microsporidia comprises >1,200 species, only 15 of which are known to infect humans, including the genera Trachipleistophora, Pleistophora, and Brachiola. We report an infection by Tubulinosema sp. in an immunosuppressed patient.
Assuntos
Hospedeiro Imunocomprometido , Microsporídios , Miosite/diagnóstico , Injúria Renal Aguda/complicações , Injúria Renal Aguda/imunologia , Idoso , Evolução Fatal , Feminino , Humanos , Leucemia Linfocítica Crônica de Células B/complicações , Leucemia Linfocítica Crônica de Células B/imunologia , Músculo Esquelético/microbiologia , Músculo Esquelético/patologia , Miosite/complicações , Miosite/microbiologiaRESUMO
We report 13 cases of Naegleria fowleri primary amebic meningoencephalitis in persons in Karachi, Pakistan, who had no history of aquatic activities. Infection likely occurred through ablution with tap water. An increase in primary amebic meningoencephalitis cases may be attributed to rising temperatures, reduced levels of chlorine in potable water, or deteriorating water distribution systems.
Assuntos
Infecções Protozoárias do Sistema Nervoso Central/epidemiologia , Doenças Transmissíveis Emergentes/epidemiologia , Naegleria fowleri/patogenicidade , Adolescente , Adulto , Amebíase/epidemiologia , Amebíase/parasitologia , Amebíase/fisiopatologia , Animais , Infecções Protozoárias do Sistema Nervoso Central/parasitologia , Infecções Protozoárias do Sistema Nervoso Central/fisiopatologia , Doenças Transmissíveis Emergentes/parasitologia , Doenças Transmissíveis Emergentes/fisiopatologia , DNA de Protozoário/análise , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Feminino , Água Doce/parasitologia , Temperatura Alta , Humanos , Masculino , Pessoa de Meia-Idade , Naegleria fowleri/genética , Naegleria fowleri/isolamento & purificação , Paquistão/epidemiologia , Reação em Cadeia da Polimerase , Abastecimento de Água , Adulto JovemRESUMO
Entamoeba histolytica is known to cause intestinal and extra-intestinal disease while the other Entamoeba species are not considered to be pathogenic. However, all Entamoeba spp. should be reported when identified in clinical samples. Entamoeba polecki, Entamoeba coli, and Entamoeba hartmanii can be differentiated morphologically from E. histolytica, but some of their diagnostic morphologic features overlap. E. histolytica, Entamoeba dispar, and Entamoeba moshkovskii are morphologically identical but can be differentiated using molecular tools. We developed a polymerase chain reaction (PCR) procedure followed by DNA sequencing of specific regions of 18S rRNA gene to differentiate the Entamoeba spp. commonly found in human stools. This approach was used to analyze 45 samples from cases evaluated for the presence of Entamoeba spp. by microscopy and a real-time PCR method capable of differential detection of E. histolytica and E. dispar. Our results demonstrated an agreement of approximately 98% (45/44) between the real-time PCR for E. histolytica and E. dispar and the 18S rRNA analysis described here. Five previously negative samples by microscopy revealed the presence of E. dispar, E. hartmanii, or E. coli DNA. In addition, we were able to detect E. hartmanii in a stool sample that had been previously reported as negative for Entamoeba spp. by microscopy. Further microscopic evaluation of this sample revealed the presence of E. hartmanii cysts, which went undetected during the first microscopic evaluation. This PCR followed by DNA sequencing will be useful to refine the diagnostic detection of Entamoeba spp. in stool and other clinical specimens.
