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Cytotechnology

, Volume 66, Issue 5, pp 861–873 | Cite as

Sensitivity of biochemical test in comparison with other methods for the detection of mycoplasma contamination in human and animal cell lines stored in the National Cell Bank of Iran

  • Vahid Molla Kazemiha
  • Amir Amanzadeh
  • Arash Memarnejadian
  • Shahram Azari
  • Mohammad Ali Shokrgozar
  • Reza Mahdian
  • Shahin Bonakdar
Original Research

Abstract

Mycoplasma contamination in cell culture is considered as serious problem in the manufacturing of biological products. Our goal in this research is to find the best standard and rapid method with high sensitivity, specificity, accuracy and predictive values of positive and negative results for detection of mycoplasma contamination in cell cultures of the National Cell Bank of Iran. In this study, 40 cell lines suspected to mycoplasma contamination were evaluated by three different methods: microbial culture, enzymatic mycoalert® and molecular. Enzymatic evaluation was performed using the mycoalert® kit while in the molecular technique, a universal primer pair was designed based on the common and fixed 16SrRNA ribosomal sequences used. Mycoplasma contaminations in cell cultures with molecular, enzymatic and microbial culture methods were determined as 57.5, 52.5 and 40 %, respectively. These results confirmed the higher rate of sensitivity, specificity and accuracy for the molecular method in comparison with enzymatic and microbial methods. Polymerase chain reaction (PCR) assay based on fixed and common sequences in the 16SrRNA, is a useful valuable and reliable technique with high sensitivity, specificity and accuracy for detection of mycoplasma contamination in cell cultures and other biological products. The enzymatic mycoalert® method can be considered as a substitution for conventional microbial culture and DNA staining fluorochrome methods due to its higher sensitivity, specificity and speed of detection (<20 min).

Keywords

Mycoalert® Cell culture Mycoplasma contamination Human-animal cell lines 

Notes

Acknowledgments

The authors would like to express their appreciation to Dr. Ehsan Mostafavi (head of Department of Epidemiology, Pasteur Institute of Iran) and also would like to thank National Cell Bank of Iran Pasteur Institute for their financial assistance.

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Vahid Molla Kazemiha
    • 1
  • Amir Amanzadeh
    • 1
  • Arash Memarnejadian
    • 2
  • Shahram Azari
    • 1
  • Mohammad Ali Shokrgozar
    • 1
  • Reza Mahdian
    • 3
  • Shahin Bonakdar
    • 1
  1. 1.National Cell Bank of IranPasteur Institute of IranTehranIran
  2. 2.Department of Hepatitis and AIDSPasteur Institute of IranTehranIran
  3. 3.Molecular Medicine Group, Department of BiotechnologyPasteur Institute of IranTehranIran

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