Multiplex PCR for identifying mycobacterial isolates.
OBJECTIVE
To develop a multiplex polymerase chain reaction (PCR) method to facilitate identification of mycobacterial isolates.


METHODS
Type strains of 14 species of mycobacteria and 56 clinical isolates were lysed by boiling in TE Triton. The lysate (5 microliters) was used directly in a PCR reaction incorporating three pairs of PCR primers expected to amplify fragments from the genome of (a) all mycobacteria, (b) Mycobacterium tuberculosis complex only and (c) M avium only. PCR products were visualised by electrophoresis on agarose gels.


RESULTS
Multiplex PCR applied to 14 type strains yielded patterns on electrophoresis which permitted identification of the mycobacterial isolates as M tuberculosis complex, M avium or as mycobacteria other than the former. The identification of 56 clinical isolates by multiplex PCR was consistent with other methods and was accomplished in less than one working day.


CONCLUSIONS
This method may facilitate rapid and convenient identification of most clinical isolates of mycobacteria by PCR and gel electrophoresis. Further evaluation is warranted.
Cormican
M
M
Department of Medical Microbiology, University College Hospital, Galway, Ireland.
Glennon
M
M
Riain
U N
UN
Flynn
J
J
D001483:Base Sequence; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000145:classification*; D009162:Mycobacterium avium / Q000145:classification; D009169:Mycobacterium tuberculosis / Q000145:classification; D016133:Polymerase Chain Reaction / Q000379:methods*