Routine diagnosis is limited to the relatively few mollicutes involved in human disease. Mycoplasmas can be diagnosed 1) indirectly by testing patient's sera for specific antibodies, 2) directly by cultivation and identification, and 3) by methods of molecular biology. Ureaplasmas are usually not diagnosed by serological means but by cultivation or PCR. Sero-diagnosis (IgM and IgG) can be achieved by complement fixation assays or enzyme-linked immunosorbent assays. For M. pneumoniae a commercial glycolipid antigen preparation is available which is suitable for a preliminary diagnosis only, as this antigen is not entirely specific. Metabolic inhibition tests or (hem)agglutination assays are now less frequently in use. Cold agglutinins, though not specific, may be an indication of an M. pneumoniae infection. Successful cultivation and isolation requires careful collection of samples from the appropriate sites, short transportation times or preservation, and the use of different formulations of broth and agar media in a specialized laboratory. All these parameters are critical, and cultivation may require 2 weeks or longer. A great number of poly- and monoclonal antibodies specific for several mycoplasma species or different serovars of U. urealyticum are available. These serve in the identification of individual colonies on agar blots or in western blot analysis. Also DNA restriction or protein separation patterns may help to identify less frequently occurring species. As myco-

plasmas quickly become nonviable during transport of clinical material and in body fluids containing antibiotic substances, strain-specific DNA probes for in situ detection and PCR have been developed for all pathogenic mycoplasmas. The sensitivity and specificity of PCR comes close to or surpasses that of cultivation methods. For all that, culture confirmation may be essential because PCR is notorious for possible false-positivity, and antimicrobial sensitivity testing and future reference cultures may be needed for clinical purposes. However, used judiciously, molecular tools will certainly change epidemiological data on mycoplasmas, as the true incidence and prevalence of these organisms becomes apparent.

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