[Antiviral action of condensed mineral polyanions. In vitro inhibition of rubella virus].
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Biomedical subjects
Publications and source records attributed to C Bonissol.
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A microtechnique is described which permits simple evaluation of the activity of the enzyme adenosine phosphorylase (AdoP), present in all mycoplasmas tested thus far. The good solubility and stability of AdoP and the sensitivity of the assay should be advantageous in detecting mycoplasmas in biological samples such as animal sera, cell cultures and vaccines.
Antisera were raised against HeLa cells and mouse cells transformed after exposure to HeLa chromosomes (ME-ch.HeLa). The antisera were positive in indirect immunofluorescence assays on both HeLa and ME-ch.HeLa cells, but were negative on normal mouse cells. Immunoprecipitation of 125I-labelled cell extracts showed that Me-ch.HeLa cells contain at their surface 3 proteins of apparent molecular weights of 185,000, 105,000 and 45,000 daltons, which were also present on the surface of our HeLa cells but not on other mouse cell lines tested. However, further study has shown that these proteins are not normal constituents of HeLa plasma membranes but are in fact surface proteins of Mycoplasma orale.
The importance of cell culture contamination by mycoplasmas is well recognized, but the means used to detect such contamination still need improvement. Most mycoplasmas possess an enzyme, adenosine phosphorylase, which is not found in cell lines. We used the ultramicromethod of Uitendaal et al. to detect the presence of mycoplasmas in sera and in tissue culture medium. The absence of adenosine phosphorylase activity seems to be the best guarantee that a serum is not contaminated by mycoplasmas. This test is also most efficient for the detection of mycoplasmas in tissue or cell cultures in vitro.
The AdoP assay has been shown to be reliable in controlling the absence of mycoplasmas in filtered sera. We therefore compared its effectiveness with that of other tests in the detection of animal cell contamination by mycoplasmas.
We have currently established the spectrum of sensitivity of the isolated strains to various antibiotics before prescribing the appropriate treatment. A microtechnic has been used. The most active antibiotics are: minocyclin, pristinamycin, erythromycin, chloramphenicol, oxytetracyclin, kanamycin, tobramycin, streptomycin, in this order.
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A total of 135 cell lines was examined for mycoplasma contamination using two techniques: isolation and specific DNA-staining with DAPI or "Hoechst 33258". The two techniques showed similar results in 64% of the cases of cell contamination while the remainder was detected only by one or the other techniques: 12,82% by the staining technique and 23% by the isolation technique, which shows that the 2 techniques are complementary. The staining technique is very quick, easy to execute, very sensitive, and should be the method of choice to detect contamination when the mycoplasma do not grow in acellular media.