Evaluation of sensitivity and specificity of a Mycoplasma haemomuris-specific polymerase chain reaction test

Abstract

Background and purpose: Mycoplasma haemomuris, a small pleomorphic bacterium parasitic of red blood cells, often causes chronic and subclinical infection of rodents. Mycoplasma haemomuris is uncultivable, and a serologic testing method is not readily available. The purpose of the study reported here was to develop a sensitive and specific polymerase chain reaction (PCR) test for detection of M. haemomuris in blood samples.

Methods: On the basis of the regions of the M. haemomuris 16S rRNA gene most divergent from corresponding regions of related bacteria, M. haemomuris-specific primers were designed so that these primers could selectively amplify M. haemomuris DNA. A PCR test was performed, using blood samples from BALB/c mice infected with M. haemomuris strains TR 8564, TR 8563, and TR 8556.

Results: Use of the PCR test enabled detection of M. haemomuris DNA in a minimum of 0.0001 microl of infected mouse blood. The test also was specific for M. haemomuris and did not amplify closely related species, such as M. haemofelis, M. haemosuis, M. orale, or Anaplasma marginale.

Conclusion: This method is sensitive and specific for detection of M. haemomuris.