Detection and confirmation of Mycoplasma pneumoniae in urogenital specimens by PCR

Abstract

Following the isolation of Mycoplasma pneumoniae from urogenital specimens (M. Goulet, R. Dular, J. G. Tully, G. Billows, and S. Kasatiya, J. Clin. Microbiol. 33:2823-2825, 1995), a study was undertaken to confirm the observations by PCR. Specific primers directed to the P1 adhesin gene of M. pneumoniae were used. A total of 300 genital specimens were tested for M. pneumoniae and Mycoplasma genitalium by culture and PCR. Of these, 15 were positive by culture and 17 were positive by PCR for M. pneumoniae. No M. genitalium was detected in any of the specimens by either method. The present study demonstrates that PCR is sensitive and rapid compared to cumbersome culture methods and can be used to detect M. pneumoniae in urogenital specimens in a routine diagnostic laboratory.

FIG. 1

(a) Agarose gel electrophoresis showing specificity of P4 primers tested with M. pneumoniae and M. genitalium. Lane 1, 100-bp ladder (Pharmacia); lane 2, DNA from M. pneumoniae ATCC 15531 amplified with P4 primers; lane 3, DNA from a clinical isolate positive for M. pneumoniae; lane 4, DNA from a clinical isolate negative for M. pneumoniae; lane 5, DNA from M. genitalium ATCC 33530 amplified with P4 primers; lane 6, 100-bp ladder (Pharmacia). (b) Southern blot of the gel shown in Fig. 1a hybridized with the probe 4A4B specific for M. pneumoniae. Lanes 2 and 3, the probe hybridized to the 345-bp M. pneumoniae-specific fragment only.

FIG. 2

(a) Agarose gel electrophoresis showing detection of M. pneumoniae in clinical specimens by PCR. Lanes 1 and 20, 100-bp ladder (Gibco BRL); lane 2, DNA from M. pneumoniae ATCC 15531 amplified with P4 primers; Lanes 3 to 19, DNA from clinical isolates amplified by P4 primers. (b) Southern blot of gel shown in panel a hybridized with probe 4A4B specific for M. pneumoniae. The probe hybridized to the 345-bp M. pneumoniae-specific fragment only in lanes 2 to 19.