Laboratory diagnosis of mycoplasmal and ureaplasmal infections

Lung Diseases - Selected State of the Art Reviews

“…Nevertheless, the complex and timeconsuming nature of many of the serological assays that have been used in the past have limited acceptance of serology for routine diagnostic testing. [Cassell et al, 1994a;Waites et al, 2000]. At present, besides research laboratories, no serologic tests assays for mycoplasmas other than M. pneumoniae have been standardized for diagnostic purposes in routine clinical microbiology laboratories nor made commercially available elsewhere.…”

“…Furthermore, there is no ideal formulation of culture media for all pathogenic species, mainly due to their different substrate and pH requirements [Waites et al, 2000]. Modified SP-4 media (broth and agar) [Lo et al, 1993], containing both glucose and arginine, can support the growth of all human pathogenic Mycoplasma species, including the fastidious M. pneumoniae and M. genitalium.…”

“…It is noteworthy that mycoplasmal etiology of respiratory diseases is considered only after failure of diagnosis of other common bacterial etiologies. In addition, there are few specialized or reference laboratories and skilled personnel [Cassell et al, 1994a;Waites et al, 2000].…”

“…Broths with any color change, and subsequent blind broth passages, should be subcultured to SP4 agar, incubated, and examined under the low-power objective of the light microscope in order to look for development of typical "fried egg"-like colonies of up to 100 µm in diameter (Figure 2). Examination of agar plates must be done on a daily basis during the first week, and thereafter every 3 to 4 days until completing 5 weeks or until growth is observed [Waites et al, 2000[Waites et al, , 2004. M. genitalium, M. fermentans and M. penetrans are also glucose-fermenting and formed colonies morphologically indistinguishable from those of M. pneumoniae, thus serologic-based definitive identification can be done by growth inhibition, metabolic inhibition, and mycoplamacidal tests [Cassel et al, 1993].…”

Current Status of the Mollicute (Mycoplasma) Lung Disease: Pathogenesis, Diagnostics, Treatment and Prevention

“…Tiny brown or black irregular colonies of Ureaplasma species develop between 1 -5 days on A8 agar plates, due to urease production in the presence of manganese sulfate (Figure 2). Typical fried egg colonies are produced by M. hominis in this medium [Cassell et al, 1994a, Waites et al, 2000.…”