Genus- and species-specific DNA probes to identify mycobacteria using the polymerase chain reaction.
Differential diagnosis of Mycobacterium tuberculosis, M. avium, and other mycobacteria remains a lengthy process. Recently, the use of DNA probes has been proposed as a new approach for a more specific and rapid diagnosis. Here, we report the cloning and sequencing of a genus-specific probe for Mycobacterium and a species-specific M. avium probe. The genus-specific probe hybridizes with DNA from nine ATCC type strains and 13 isolates of mycobacteria but not to non-mycobacterial DNA. In addition, the cloned fragment could also be amplified by polymerase chain reaction (PCR) in DNa of ten different mycobacterial type strains. The M. avium specific probe hybridizes strongly to sequences amplified in M. avium but not other mycobacterial or non-mycobacterial DNA. Amplification of the target sequence by PCR allowed the detection of 1 fg of all mycobacterial DNA tested for the genus-specific probe and 1 fg of M. avium DNA for the species-specific probe.
Fries
J W
JW
Department of Tropical Public Health, Harvard School of Public Health, Boston, Massachusetts 02115.
Patel
R J
RJ
Piessens
W F
WF
Wirth
D F
DF
D000818:Animals; D001483:Base Sequence; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D005784:Gene Amplification; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics* / Q000302:isolation & purification; D009162:Mycobacterium avium / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D013045:Species Specificity
D015342:DNA Probes; D004269:DNA, Bacterial