Rapid Detection of Mycoplasma-Infected Cells by an ssDNA Aptamer Probe

Yanting Liu, Wenqi Jiang, Shuanghui Yang, Jianzhong Hu, Hongbin Lu, Wei Han, Jianguo Wen, Zihua Zeng, Jianjun Qi, Ling Xu, Haijun Zhou, Hongguang Sun, Youli Zu

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Mycoplasmas are unique cell wall-free bacteria. Because they lack a cell wall and have resistance to β-lactam antibiotics, mycoplasma is the major pathogen that infects cultured cells in research laboratories. For rapid detection of mycoplasma-infected cells, we developed an ssDNA aptamer sequence composed of 40 nucleotides. Flow cytometry analysis showed that the synthetic aptamer probe selectively targeted mycoplasma-infected culture cells with high specificity identical to commercially available PCR-based assays. Additionally, fluorescent microscopy studies revealed that the aptamer probe rapidly stained mycoplasma-infected cells with higher sensitivity compared to Hoechst dye-mediated cellular DNA content stains. Moreover, confocal microscopy studies of trypsin-treated cells validated that the aptamer probes selectively targeted mycoplasma components on the surface of infected cells. Finally, preclinical studies of peripheral blood cells demonstrated that the aptamer probe was able to detect in vitro mycoplasma infection of primary lymphocytes. Taken together, these findings indicate that the aptamer probe will not only allow rapid detection of mycoplasma-infected culture cells for research purposes but also provide a simple method to monitor mycoplasma infection in primary cell products for clinical use.

Original languageEnglish (US)
Pages (from-to)2028-2038
Number of pages11
JournalACS Sensors
Volume4
Issue number8
DOIs
StatePublished - Aug 23 2019

Keywords

  • aptamer probe
  • cell culture
  • mycoplasma infection
  • primary lymphocytes
  • rapid detection

ASJC Scopus subject areas

  • Bioengineering
  • Instrumentation
  • Process Chemistry and Technology
  • Fluid Flow and Transfer Processes

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