Using oligonucleotide suspension arrays for laboratory identification of bacteria responsible for bacteremia
The aim of this study was to develop and validate an oligonucleotide suspension array for rapid identification of 15 bacterial species responsible for bacteremia, particularly prevalent in Chinese hospitals. The multiplexed array, based on the QIAGEN LiquiChip Workstation, included 15 oligonucleotide probes which were covalently bound to different bead sets. PCR amplicons of a variable region of the bacterial 23S rRNA genes were hybridized to the bead-bound probes. Thirty-eight strains belonging to 15 species were correctly identified on the basis of their corresponding species-specific hybridization profiles. The results show that the suspension array, in a single assay, can differentiate isolates over a wide range of strains and species, and suggest the potential utility of suspension array system to clinical laboratory diagnosis.
Key wordsOligonucleotide array Bacteremia 23S rRNA Multiplexed detection
Unable to display preview. Download preview PDF.
- Bovers, M., Diaz, M.R., Hagen, F., Spanjaard, L., Duim, B., Visser, C.E., Hoogveld, H.L., Scharringa, J., Hoepelman, I.M., Fell, J.W., Boekhout, T., 2007. Identification of genotypically diverse Cryptococcus neoformans and Cryptococcus gattii isolates by Luminex xMAP technology. J. Clin. Microbiol., 45(6):1874–1883. [doi:10.1128/JCM.00223-07]PubMedCrossRefGoogle Scholar
- Ding, L.P., Sun, Y.H., Wang, Q., Nian, H., 2004. Drug resistance of common bacteria isolated from blood and bone marrow. J. Chin. Med. Univ., 33(1):83–85 (in Chinese).Google Scholar
- Dunbar, S.A., Vander Zee, C.A., Oliver, K.G., Karem, K.L., Jacobson, J.W., 2003b. Quantitative, multiplexed detection of bacterial pathogens: DNA and protein applications of the Luminex LabMAP system. J. Microbiol. Methods, 53(2):245–252. [doi:10.1016/S0167-7012(03)00028-9]PubMedCrossRefGoogle Scholar
- Leffers, H., Kjems, J., Ostergaard, L., Larsen, N., Garrett, R.A., 1987. Evolutionary relationships amongst archaebacteria. A comparative study of 23S ribosomal RNAs of a sulphur-dependent extreme thermophile, an extreme halophile and a thermophilic methanogen. J. Mol. Biol., 195(1):43–61. [doi:10.1016/0022-2836(87)90326-3]PubMedCrossRefGoogle Scholar
- Maiwald, M., Ditton, H.J., Sonntag, H.G., von Knebel Doeberitz, M., 1994. Characterization of contaminating DNA in Taq polymerase which occurs during amplification with a primer set for Legionella 5S ribosomal RNA. Mol. Cell. Probes, 8(1):11–14. [doi:10.1006/mcpr.1994.1002]PubMedCrossRefGoogle Scholar
- Nolan, J.P., Mandy, F.F., 2001. Suspension array technology: new tools for gene and protein analysis. Cell. Mol. Biol. (Noisy-le-grand), 47(7):1241–1256.Google Scholar