Identification of Mycobacterium intracellulare by a polymerase chain reaction using species-specific primers.
METHODS
The polymerase chain reaction (PCR) is a rapid and specific method used to amplify a certain DNA fragment. It is applicable to rapid diagnosis of mycobacterial infections. By use of species-specific primers, it is possible to identify mycobacteria by PCR. In this study, a newly constructed primer was tested for specificity for Mycobacterium intracellulare in the PCR.


OBJECTIVE
M. intracellulare is one of the most frequently found bacteria in opportunistic infection in AIDS, and rapid identification of this species is important. The purpose of this study was to construct a primer specific to this species as a suitable tool for identification.


METHODS
PCR products of M. tuberculosis and M. intracellulare, obtained by using the primers YNP-1 and YNP-2, were sequenced and compared. They showed a difference in the base sequences. A sequence unique to M. intracellulare was used as the primer specific to this species. Various mycobacterial and non-mycobacterial DNAs were used as the primer specific to this species. Various mycobacterial and non-mycobacterial DNAs were used as the template to evaluate the specificity of the newly constructed primers, YNP-7 and YNP-8. Sputum samples were also examined by PCR using the primers.


RESULTS
In total 25 species of culture mycobacterial and non-mycobacterial strains and 76 sputum samples were tested by PCR. Only M. intracellulare DNA was amplified with PCR using the primers YNP-7/8.


CONCLUSIONS
The specificity of the newly constructed primers for M. intracellulare was confirmed.
Yamazaki
T
T
Department of Bacteriology, National Institute of Health, Tokyo, Japan.
Nakamura
R M
RM
D015373:Bacterial Typing Techniques; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000145:classification; D015269:Mycobacterium avium Complex / Q000302:isolation & purification*; D015270:Mycobacterium avium-intracellulare Infection / Q000175:diagnosis; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D013183:Sputum / Q000382:microbiology
D017931:DNA Primers; D004269:DNA, Bacterial