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Cytotechnology

, Volume 61, Issue 3, pp 117–124 | Cite as

PCR-based detection and eradication of mycoplasmal infections from various mammalian cell lines: a local experience

  • Vahid Molla Kazemiha
  • Mohammad Ali Shokrgozar
  • Mohammad Reza Arabestani
  • Morteza Shojaei Moghadam
  • Shahram Azari
  • Susan Maleki
  • Amir Amanzadeh
  • Mahmood Jeddi Tehrani
  • Fazel ShokriEmail author
Original Research

Abstract

A total of 200 cell lines including different human, monkey, mice, hamster and rat cell types were examined for mycoplasma infection status. PCR assay using generic-specific universal primers showed that 40 (20%) of the cell lines are contaminated with mycoplasma. Employment of species-specific primers within these infected cell lines revealed infection with M. hyorhinis (42.5%), M. fermentas (37.5%), M. arginini (37.5%), M. orale (12.5%) and A. laidlawii (7.5%). A number of the cultures were coinfected with 2 or 3 different species. Contaminated samples were treated with BM-Cyclin, Ciprofloxacin and mycoplasma removal agent (MRA). Mycoplasma eradication was subsequently checked by PCR following 2 weeks continuous culture of treated cells in antibiotic free culture medium. Mycoplasmal infections were eradicated in 100, 70 and 42% of infected cell lines when the samples were treated with BM-Cyclin, MRA and Ciprofloxacin, respectively. However, 12% (BM-Cyclin), 62.5% (MRA) and 82.5% (Ciprofloxacin) of mycoplasma regrowth was observed 4 months after the treatment. Notably, the risk of spontaneous culture death was 17.5, 12.5 and 0% for BM-Cyclin, MRA and Ciprofloxacin, respectively.

Keywords

Mycoplasmal infection Mammalian cell lines Elimination Antibiotics PCR-based detection 

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Vahid Molla Kazemiha
    • 1
  • Mohammad Ali Shokrgozar
    • 1
  • Mohammad Reza Arabestani
    • 1
  • Morteza Shojaei Moghadam
    • 2
  • Shahram Azari
    • 1
  • Susan Maleki
    • 3
  • Amir Amanzadeh
    • 1
  • Mahmood Jeddi Tehrani
    • 1
    • 4
    • 5
  • Fazel Shokri
    • 1
    • 6
    Email author
  1. 1.National Cell Bank of Iran, Pasteur Institute of IranTehranIran
  2. 2.Faculty of Biotechnology and Biomolecular SciencesUniversiti Putra MalaysiaUPM SerdangMalaysia
  3. 3.Department of Biology, School of ScienceShahid Chamran UniversityAhvazIran
  4. 4.Monoclonal Antibody Research CenterAvicenna Research InstituteTehranIran
  5. 5.Immune and Gene Therapy Laboratory, Cancer Center KarolinskaKarolinska InstituteStockholmSweden
  6. 6.Department of Immunology, School of Public HealthTehran University of Medical SciencesTehranIran

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