An ultramicrochemical test for mycoplasmal contamination of cultured cells |
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Authors: | M P Uitendaal C H M M De Bruyn M Hatanaka P Hösli |
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Institution: | (1) Department of Molecular Biology, Institut Pasteur, 25, Rue du Dr. Roux, 75015 Paris, France;(2) Present address: Department of Human Genetics, Faculty of Medicine, University of Nijmegen, Nijmegan, The Netherlands;(3) Present address: Laboratory of DNA Tumor Viruses, National Cancer Institute, National Institutes of Health, Bethesda, Maryland |
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Abstract: | Summary A sensitive ultramicrochemical enzyme test for mycoplasmal contamination of cultured cells, based on the determination of
the activity of adenosine phosphorylase, is described. The test was performed by assaying the enzymatic conversion of 8-14C]adenine and ribose-1-phosphate to 8-14C]adenosine by incubating a plastic leaflet carrying a counted number of cells (1 to 10). These leaflets were isolated from
the bottom of the same plastic film dish in which the cells were cultured for experimental or diagnostic purposes, e.g. prenatal
diagnosis or inborn errors of metabolism. The present test should be several 1000-fold more sensitive than the originally
reported enzymatic method because (a) the adenosinephosphorylase reaction is measured in the nucleoside forming direction
which is by far the most active; and (b) the assay is performed with the cells and not with the culture medium. The latter
is of special importance for the detection of those low-grade contaminations in which most of the mycoplasma particles are
attached to cell membranes.
This investigation was partly supported by FUNGO (The Netherlands), INSERM (France) (A. T. P. 36.76.68), and DGRST (France)
(No. 75.50.004). |
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Keywords: | mycoplasma adenosine phosphorylase ultramicrochemical enzyme tests cell cultures |
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