Parallel Detection of Bacterial Pathogens in Cerebrospinal Fluid with 16S rRNA Probe Microarray

摘要

To establish the microarray technique with 16S rRNA probe for the rapid detection of pathogenic bacteria in cerebrospinal fluid, a set of Digoxin labelling universal primers were designed, and used to amplify the 16S rRNA gene, and 2 specific probes for each bacterium designed were spotted onto nylon membrane to make a microarray, followed by hybridization to PCR products of bacteria, which set up a microarray technique for parallel detection of bacteria in cerebrospinal fluid (CSF). Twenty-nine CSF specimens from 16 patients that were proved to be positive by bacterial cultures and by using Vitek AMA-32 automated system were examined in the present investigation. It was found that positive fragment with length of 168?bp were amplified from 45 strains of 20 bacterial species, and amplifications also detected in 29 CSF specimens. The method of universal PCR could detect as few as 1 pg of bacterial DNA, and this microarray could identify all the bacterial species in CSF except Mycobacterium tuberculosis and Proteus mirabilis , satisfying the direct detection of bacteria in CSF specimens including a case of mixed bacterial infection. The positive and negative predictive value as well as the diagnostic index of this microarray were 100%, 94.67%, and 180.88 respectively, and the time of performance spent only for 5?h. It was concluded the method described in the present investigation can be used to detect and identify the pathogenic bacteria in CSF.