Development and evaluation of 16S rDNA microarray for detecting bacterial pathogens in cerebrospinal fluid.
The rapid identification of bacteria in cerebrospinal fluid (CSF) is very important for patient management and antimicrobial therapies. We developed a 16S DNA microarray-based method that targets 16S rDNA and can directly detect bacteria from CSF without cultivation. Universal primers and specific probes were designed from the 16S rDNA sequence data retrieved directly from the GenBank database. The specificity of the assay is obtained through a combination of microarray hybridization and enzymatic labeling of the constructed specific probes. Cultivation-dependent assays were used as reference methods in the development and evaluation of the method. With the exception of Mycobacterium tuberculosis and Proteus mirabilis, forty-five positive blood culture media were successfully differentiated. When this procedure was applied directly to 100 CSF specimens, 29 specimens from 16 patients were positive by bacterial culture and 3 culture-positive CSF specimens produced no hybridized signals. The remaining 26 specimens were correctly identified, including one with mixed infection. The accuracy, sensitivity, and specificity of the assay can be increased further by designing more oligonucleotides for the microarray. This method is versatile and makes it possible to detect more bacteria in a single assay and discriminate different bacterial genera.
Liu
Yi
Y
Key Laboratory for Biotech-Drugs Ministry of Health, Shandong Medicinal Biotechnology Center, Jinan 250062, China.
Han
Jin-Xiang
JX
Huang
Hai-Yan
HY
Zhu
Bo
B
D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification; D001424:Bacterial Infections / Q000175:diagnosis*; D001483:Base Sequence; D002555:Cerebrospinal Fluid / Q000382:microbiology*; D017931:DNA Primers; D004275:DNA, Ribosomal / Q000235:genetics*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S / Q000235:genetics*
D017931:DNA Primers; D004275:DNA, Ribosomal; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S