Oligonucleotide microarray for identification of Bacillus anthracis based on intergenic transcribed spacers in ribosomal DNA.
We developed a DNA microarray for identification of Bacillus anthracis and other phylogenetic groupings within the "Bacillus cereus group". Nucleotide sequences of 16S-23S ribosomal DNA internal transcribed spacers containing genes for tRNA(Ile) from 52 B. anthracis strains were found to be identical to sequences from seven strains published previously and different from all other bacteria. When 42 oligonucleotide probes targeting polymorphic sites were immobilized on glass slides and hybridized to fluorescently labeled PCR amplification products, one or more mismatches could be discriminated in all but one cases. Hence, hybridization events were highly specific and identification of B. anthracis was straightforward.
Nübel
Ulrich
U
Robert Koch Institut, 13353 Berlin, Germany. nuebelu@rki.de
Schmidt
Peter M
PM
Reiss
Edda
E
Bier
Frank
F
Beyer
Wolfgang
W
Naumann
Dieter
D
D001408:Bacillus anthracis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D020137:Base Pair Mismatch; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D010802:Phylogeny; D012329:RNA, Bacterial / Q000235:genetics; D012355:RNA, Transfer, Ile / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer; D012329:RNA, Bacterial; D012355:RNA, Transfer, Ile