Towards accurate exclusion of neonatal bacterial meningitis: a feasibility study of a novel 16S rDNA PCR assay

摘要

PCRctic: a CSF samples; b Intact bacteria are pelleted along with patient cells and debris; c The pellet is resuspended in the assay mixture (PCR reagents, ethidium azide, buffer); d light activates the ethidium azide, destroying exposed DNA (from dead bacteria or from contaminants in the reagents) but leaving intact bacteria unaffected; during PCR e, heat causes the bacteria to lyse, making their DNA available for amplification. f amplification products are then detected by a simple fluorometric assay (melting-curve analysis) using a widely-available real-time PCR machine. Total assay time is approximately 2 h