Detection of Mycoplasma pulmonis in experimentally infected laboratory rats by 16S rRNA amplification.
Recently, an rRNA-based polymerase chain reaction (PCR) has been developed for the detection of murine mycoplasmas at both the genus and species level (F. J. M. van Kuppeveld, J. T. M. van der Logt, A. F. Angulo, M. J. van Zoest, W. G. V. Quint, H. G. Niesters, J. M. D. Galama, and W. J. G. Melchers, Appl. Environ. Microbiol. 58:2606-2615, 1992). In this study, the diagnostic value of this PCR assay for the detection of Mycoplasma pulmonis in infected rats was studied. For this purpose, 25 Wistar rats were infected intranasally with M. pulmonis strain M72-138 and investigated for the presence of this pathogen by both in vitro isolation and PCR. Five rats were monitored longitudinally by screening of throat swabs at several time points for up to 248 days postinfection. The remaining 20 rats were killed between 3 and 87 days postinfection, and organism recovery from both throat and urogenital tract specimens was attempted. M. pulmonis could be detected in the throat for up to 248 days postinfection but not in the urogenital tract, either by culture or by PCR. PCR proved to be the optimal method for testing throat samples. All samples in which M. pulmonis was detected by culture were also positive by PCR. By PCR, M. pulmonis was also detected in 3.7% of the samples which were culture negative and in 9.9% of the samples from which cultures were overgrown with bacteria. The results of this study demonstrate the suitability of PCR for the detection of mycoplasmal infection in rodents.
van Kuppeveld
F J
FJ
Department of Medical Microbiology, University of Nijmegen, The Netherlands.
Melchers
W J
WJ
Willemse
H F
HF
Kissing
J
J
Galama
J M
JM
van der Logt
J T
JT
D000818:Animals; D001483:Base Sequence; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000235:genetics / Q000382:microbiology*; D010614:Pharynx / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains; D017208:Rats, Wistar; D014566:Urogenital System / Q000382:microbiology
D012336:RNA, Ribosomal, 16S