Document Detail

Indicator cell lines for the detection of hidden mycoplasma contamination, using an adenosine phosphorylase screening test.
MedLine Citation:
PMID:  3141514     Owner:  NLM     Status:  MEDLINE    
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.
G T Spierenburg; A A Polak-Vogelzang; B J Bast
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of immunological methods     Volume:  114     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  1988 Nov 
Date Detail:
Created Date:  1988-12-19     Completed Date:  1988-12-19     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  115-9     Citation Subset:  IM    
Department of Clinical Immunology, University Hospital, Utrecht, The Netherlands.
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MeSH Terms
Biological Markers / analysis*
Cell Line
Cell-Free System
Fibroblasts / enzymology*,  microbiology
Mycoplasma / enzymology*
Pentosyltransferases / metabolism*
Purine-Nucleoside Phosphorylase / metabolism*
Reg. No./Substance:
0/Biological Markers; EC 2.4.2.-/Pentosyltransferases; EC 2.4.2.-/adenosine phosphorylase; EC Phosphorylase

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