Abstract
In the present report we describe a rapid ans sensitive assay for mycoplasma detection in cell cultures. The assay is based on the ability of contaminated culture supernatants to modulate [ 3H]TdR incorporation by unstimulated mouse splenocytes. Several mycoplasma species ( Mycoplasma orale, culturable and non-culturable strains of Mycoplasma hyorhinis) inhibited [ 3H]TdR incorporation and permitted the detection of some contaminated cell cultures that would otherwise have escaped detection in assays measuring [ 3H]TdR incorporation by mitogen-stimulated splenocytes. On the other hand, several other mycoplasma species ( Mycoplasma arginini, Mycoplasma hominis) strongly enhanced [ 3H]TdR incorporation by unstimulated splenocytes. This enhancement was optimally detectable on day 2 after initiation of the cultures. The sensitivity of the assay was determined for a mycoplasma species (culturable M. hyorhinis) that inhibited as well as for one ( M. arginini) that enhanced [ 3H]TdR incorporation. In both cases, the sensitivity was such that 1−3× 10 2 mycoplasma colony-forming units (CFU) could be detected.
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