16S rRNA-Targeted identification of cyanobacterial genera usingoligonucleotide-probes immobilized on bacterial magnetic particles

摘要

16S rRNA-targeted identification of cyanobacterial genera, Anabaena, Microcystis, Nostoc, Oscillatoria, Synechococcus was developed using bacterial magnetic particles (BMPs). 16S rRNA-targeted capture probes designed from the genus specific region of the 16S rRNA sequence were immobilized on BMPs. Identification of cyanobacteria was per-formed by a sandwich hybridization using the capture probes-BMP conjugates and a digoxigenin (DIG)-labeled detector probe complementary to the highly conserved 16S rRNA sequence for cyanobacteria. The luminescence intensity of the probe/target-BMP hybrids was measured after reaction with alkaline phosphatase conjugated anti-DIG antibody. Five species of cyanobacteria from five different genera were successfully discriminated using this magnetic capture system.