Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Biotechnical Methods Section (BTS)

Detection of mycoplasma in leukemia–lymphoma cell lines using polymerase chain reaction


The specific, sensitive and reliable detection of mycoplasma contamination in cell cultures is an important part of mycoplasma control. We have sought to develop and validate a method for mycoplasma detection which is sensitive and accurate, but also practical in the sense of time spent, costs, and applicability in the standard laboratory; finally, the method should be suitable for screening large numbers of test specimens. To that end, we adapted a previously developed polymerase chain reaction (PCR) method for daily routine application. This single-step PCR uses a mixture of primers annealing to gene sequences coding for evolutionarily conserved 16S rRNA of different mycoplasma species, including the ones most commonly found in cell cultures. An internal control was introduced to exclude any false-negative tests resulting from technical PCR problems. This mycoplasma detection by PCR has been validated prospectively on 201 consecutive leukemia–lymphoma cell lines received at the institute over a 3-year period and on 118 initially positive cell lines after anti-mycoplasma treatment with antibiotics. The sensitivity (detection of true positives) of this PCR detection assay was 96% and the specificity (detection of true negatives) was also 96%, with positive and negative predictive values (probability of correct result) of 86% and 99%, respectively. PCR defined the mycoplasma status with 96% accuracy (detection of true positives and true negatives). Besides the high sensitivity and specificity, further attractive features of the PCR approach are the ease and speed with which large numbers of specimens can be tested. PCR mycoplasma analysis provides a readily available, quick and reliable test system with which to manage the important issue of mycoplasma contamination of cell lines.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it


Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2


  1. Drexler HG, Uphoff CC . Contamination of cell culture, mycoplasma. In: Spier E et al (eds) The Encyclopedia of Cell Technology Wiley: New York 2000 pp 609–627

    Google Scholar 

  2. Uphoff CC, Brauer S, Grunicke D, Gignac SM, MacLeod RAF, Quentmeier H, Steube K, Tümmler M, Voges M, Wagner B, Drexler HG . Sensitivity and specificity of five different mycoplasma detection assays Leukemia 1992 6: 335–341

    CAS  PubMed  Google Scholar 

  3. Uphoff CC, Gignac SM, Drexler HG . Mycoplasma contamination in human leukemia cell lines. I. Comparison of various detection methods J Immunol Methods 1992 149: 43–53

    Article  CAS  Google Scholar 

  4. Razin S . DNA probes and PCR in diagnosis of mycoplasma infections Mol Cell Probes 1994 8: 497–511

    Article  CAS  Google Scholar 

  5. Drexler HG (ed). The Leukemia–Lymphoma Cell Line FactsBook Academic Press: San Diego 2000

    Google Scholar 

  6. Drexler HG, Dirks WG, MacLeod RAF, Quentmeier H, Steube KG, Uphoff CC (eds). DSMZ Catalogue of Human and Animal Cell Lines, 8th edn Braunschweig, Germany 2001

    Google Scholar 

  7. Uphoff CC, Meyer C, Drexler HG . Elimination of mycoplasma from leukemia-lymphoma cell lines using antibiotics Leukemia 2002 16: 284–288

    Article  CAS  Google Scholar 

  8. Hopert A, Uphoff CC, Wirth M, Hauser H, Drexler HG . Specificity and sensitivity of polymerase chain reaction (PCR) in comparison with other methods for the detection of mycoplasma contamination in cell lines J Immunol Methods 1993 164: 91–100

    Article  CAS  Google Scholar 

  9. Wirth M, Berthold E, Grashoff M, Pfutzner H, Schubert U, Hauser H . Detection of mycoplasma contaminations by the polymerase chain reaction Cytotechnology 1994 16: 67–77

    Article  CAS  Google Scholar 

  10. Uphoff CC, Hu ZB, Gignac SM, Ma W, Rainey FA, Kreutz M, Ludwig WD, Drexler HG . Characterization of the monocyte-specific esterase (MSE) gene Leukemia 1994 8: 1510–1526

    CAS  PubMed  Google Scholar 

  11. Uphoff CC, Drexler HG . Detection of mycoplasma contaminations in cell cultures by PCR analysis Human Cell 1999 12: 229–236

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations


Rights and permissions

Reprints and Permissions

About this article

Cite this article

Uphoff, C., Drexler, H. Detection of mycoplasma in leukemia–lymphoma cell lines using polymerase chain reaction. Leukemia 16, 289–293 (2002).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


  • cell lines
  • leukemia
  • lymphoma
  • mycoplasma detection
  • PCR

This article is cited by


Quick links