| Indicator cell lines for the detection of hidden mycoplasma contamination, using an adenosine phosphorylase screening test. | |
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MedLine Citation:
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PMID: 3141514 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Mycoplasmas are a major cause of cell culture contamination and are especially troublesome during HAT selection. The enzyme adenosine phosphorylase (adoP) is present in all common mycoplasma species but is considered to have a low activity in mammalian cells. However, using an adoP screening test, we have observed that some cell cultures do possess an intrinsic adoP activity leading to false positive results. Moreover, as a false negative result, we encountered a variant of Mycoplasma orale (identified after cultivation on agar and immunostaining) which was not detectable with the adoP screening in cell culture supernatants and only at low levels in cell lysates. To increase the low signal/noise adoP ratio found there, we used an indicator cell line with low intrinsic activity. Indicator cells were inoculated with the test supernatant and the adoP activity of these infected cells were measured after lysis. The procedure diminished the effect of biological variation in intrinsic enzyme activity between the several cell lines tested. Furthermore, in another mycoplasma infected cell line (with M. fermentans), this infection was only reliably detected using these indicator cells. With this procedure we obtained rapid results which were concordant with those obtained using the time consuming cultivation on agar. |
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Authors:
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G T Spierenburg; A A Polak-Vogelzang; B J Bast |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of immunological methods Volume: 114 ISSN: 0022-1759 ISO Abbreviation: J. Immunol. Methods Publication Date: 1988 Nov |
Date Detail:
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Created Date: 1988-12-19 Completed Date: 1988-12-19 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 1305440 Medline TA: J Immunol Methods Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 115-9 Citation Subset: IM |
Affiliation:
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Department of Clinical Immunology, University Hospital, Utrecht, The Netherlands. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Biological Markers / analysis* Cell Line Cell-Free System Fibroblasts / enzymology*, microbiology Mice Mycoplasma / enzymology* Pentosyltransferases / metabolism* Purine-Nucleoside Phosphorylase / metabolism* |
| Chemical | |
Reg. No./Substance:
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0/Biological Markers; EC 2.4.2.-/Pentosyltransferases; EC 2.4.2.-/adenosine phosphorylase; EC 2.4.2.1/Purine-Nucleoside Phosphorylase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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