RESUMEN
Next-generation DNA sequencing can be used to catalog individual organisms within complex, polymicrobial specimens. Here, we utilized deep sequencing of 16S rRNA to implicate Actinomadura madurae as the cause of mycetoma in a diabetic patient when culture and conventional molecular methods were overwhelmed by overgrowth of other organisms.
Asunto(s)
Actinomycetales/aislamiento & purificación , Micetoma/diagnóstico , Actinomycetales/clasificación , Actinomycetales/genética , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Complicaciones de la Diabetes , Femenino , Pie/patología , Secuenciación de Nucleótidos de Alto Rendimiento , Histocitoquímica , Humanos , Microscopía , Persona de Mediana Edad , Datos de Secuencia Molecular , Micetoma/microbiología , Filogenia , ARN Ribosómico 16S/genéticaRESUMEN
Pythium insidiosum is an emerging human pathogen classified among brown algae and diatoms that can cause significant morbidity and mortality in otherwise healthy individuals. Here we describe a pediatric patient with pythiosis acquired in the southern United States, diagnosed by molecular screening and DNA sequencing of internal transcribed spacer region 1.
Asunto(s)
Pierna/parasitología , Reacción en Cadena de la Polimerasa , Pitiosis/diagnóstico , Pythium , Adolescente , Amputación Quirúrgica , Secuencia de Bases , Femenino , Humanos , Pierna/cirugía , Técnicas de Diagnóstico Molecular , Datos de Secuencia Molecular , Tipificación Molecular , Pitiosis/parasitología , Análisis de Secuencia de ADNRESUMEN
Strongyloides stercoralis is an important human parasite, especially in rural areas and developing countries. Infected immunosuppressed patients are at risk for hyperinfection, with severe clinical consequences. Here we describe the incidental detection and diagnosis of an unexpected S. stercoralis infection by methods designed to detect fungal 28S ribosomal DNA.
Asunto(s)
Neoplasias Hematológicas/complicaciones , Strongyloides stercoralis/aislamiento & purificación , Estrongiloidiasis/diagnóstico , Estrongiloidiasis/patología , Animales , Análisis por Conglomerados , ADN de Helmintos/química , ADN de Helmintos/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Humanos , Masculino , Microscopía , Persona de Mediana Edad , Filogenia , Reacción en Cadena de la Polimerasa , ARN Ribosómico 28S/genética , Análisis de Secuencia de ADN , Strongyloides stercoralis/genética , Estrongiloidiasis/parasitologíaRESUMEN
The absence of analytical controls for polymerase chain reaction (PCR)-based diagnostic tests for Bordetella pertussis limits their clinical utility. In this study, multiplex PCR simultaneously targeted two specific Bordetella pertussis sequences, the chromosomal repeated insertion sequence IS481 (IS) and the pertussis toxin promoter region (PT). A multi-target hybridization-EIA (Hyb-EIA) method in a 96-well microtiter-plate format was used to detect amplicons. Forty-seven (15%) of the 318 nasopharygeal specimens tested positive for at least one DNA target of B. pertussis by PCR, including the 10 known positive samples by culture and/or direct fluorescent antibody (DFA). Forty-six of the 47 PCR positive samples were considered positive for B. pertussis using the consensus interpretation criteria. Simultaneous detection of multiple chromosomal regions may identify false-positive and -negative results due to analytical variations or potential sequence polymorphism, and uncover a wider range of pathogenic strains.