Detection and identification of mycobacteria by amplification of rRNA.
Oligonucleotides specific at a genus, group, or species level were defined by a systematic comparison of small-subunit rRNA sequences from Mycobacterium tuberculosis, M. bovis, M. africanum, M. bovis BCG, M. avium, M. kansasii, M. marinum, M. gastri, M. chelonae, M. smegmatis, M. terrae, M. nonchromogenicum, M. xenopi, M. malmoense, M. szulgai, M. scrofulaceum, M. fortuitum, M. gordonae, M. intracellulare, M. simiae, M. flavescens, M. paratuberculosis, M. sphagni, M. cookii, M. komossense, M. phlei, and M. farcinica. On the basis of the defined oligonucleotides, the polymerase chain reaction technique was explored to develop a sensitive taxon-specific detection system for mycobacteria. By using M. tuberculosis as a model system, fewer than 10 bacteria could be reliably detected by this kind of assay. These results suggest that amplification of rRNA sequences by the polymerase chain reaction may provide a highly sensitive and specific tool for the direct detection of microorganisms without the need for prior cultivation.
Böddinghaus
B
B
Institut für Medizinische Mikrobiologie, Medizinische Hochschule Hannover, Federal Republic of Germany.
Rogall
T
T
Flohr
T
T
Blöcker
H
H
Böttger
E C
EC
D001483:Base Sequence; D004269:DNA, Bacterial / Q000302:isolation & purification; D005784:Gene Amplification; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000145:classification / Q000235:genetics*; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D012335:RNA, Ribosomal / Q000096:biosynthesis*; D012336:RNA, Ribosomal, 16S / Q000096:biosynthesis*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S