A general method to generate DNA probes for microorganisms.
We present a method that permits the rapid generation of DNA probes for eubacteria. In the procedure the variable regions for the 16s rRNA genes are amplified using polymerase chain reaction (PCR) technology and primers based on the conserved regions of these genes. Following sequencing of the variable regions, a choice is possible for a probe specific for that organism. No knowledge of the molecular biology of the microorganism is required prior to the application of this approach. The generality of the method is shown using Salmonella typhimurium, Staphylococcus aureus, Clostridium perfringens, Klebsiella pneumoniae, Pseudomonas fluorescens, Aeromonas salmonicida and Mycobacterium bovis. A. salmonicida was examined in detail and a DNA probe was prepared that distinguishes it from other Aeromonas species.
National Diagnostics Centre-BioResearch Ireland, University College Galway.
T
T
Barry
R
R
Powell
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F
Gannon
D000333:Aeromonas / Q000235:genetics*; D001483:Base Sequence; D015342:DNA Probes; D005784:Gene Amplification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000032:analysis*; D012335:RNA, Ribosomal / Q000032:analysis*; D012336:RNA, Ribosomal, 16S / Q000032:analysis*
D015342:DNA Probes; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S