Oligo-chips for Detecting Pathogens in Cerebrospinal Fluids:A Preliminary Study / 中华医院感染学杂志

ABSTRACT

OBJECTIVE To develop a macroarray method to detect pathogens in cerebrospinal fluid.METHODS According to the bacterial 16S rRNA genes,designed 10 kinds of specific probes and a pair of universal primers that can amplify rRNA gene of all bacteria.The tailed probes were spotted onto a nylon membrane.DNA was isolated from each pathogen,and subjected to UP-PCR to amplify target fragments,which were labeled with bio-16-dUTP at the same time.All those denatured fragments were hybridized to the probes on nylon membrane and visualized by AKP labeled avidin.The sensitivity and specificity of the system were detected.A total of 32 CSF samples,which were verified the bacterial infection by the routine method,were tested by this method.RESULTS It was sensitive to 10 CFU/ml when detecting Escherichia coli.Every kind of pathogens only reacted to its corresponding probes fixed on nylon membranes,which showed high specificity.The result of identifying 32 CSF clinical specimens accorded with that of routine method.CONCLUSIONS The method can screen out common pathogens in CSF sensitively and exactly.