Objective: To detect the rate of Mycoplasma infection in cell lines and further determine its types.

Methods: We performed nest PCR amplification of Mycoplasma's conserved regions (16S-23S) and sequenced the spacer with different length between conserved regions.

Results: Within the tested 22 cell lines, 17 (77.3%) showed Mycoplasma infections, of which 5 had two or more types of Mycoplasma. M. fermentans and hyorhinis were more frequently detected within the types of infected Mycoplasma.

Conclusion: The high rate of Mycoplasma infection in cell lines makes it necessary for researchers to pay more attention to its influence on research data when using cell lines as models. Establishment of detection and classifying techniques make it possible to further study the pathogenesis of different types of Mycoplasma.

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