Establishment of multiplex real-time PCR assay to detect AIDS-associated Mycoplasmas

Objective To establish a rapid, sensitive and specific multiplex real-time PCR assay for the detection of AIDS-associated Mycoplasmas. Methods Three sets of primers and probes were designed based on the specific sequence of the ftsZ gene (M. penetrans and M. fermentans) and the rpoB gene (M. pirum) and the Multiplex real-time PCR assay was established. Eight other Mycoplasmas and 14 pathogens were used to evaluate the specificity of the assay. The sensitivity of the assay was evaluated and compared with conventional PCR assay by using standard concentration positive plasmids. Results No specific amplifications were presented by using 22 other pathogens. The sensitivity of this Multiplex real-time PCR assay was about 10 times higher than that of conventional PCR for M. penetrans and M. fermentans, and 100 times higher than M. pirum. Conclusion This Multiplex real-time PCR assay was sensitive and specific, which could be used for the detection of AIDS-associated mycoplasmas and might be used in the clinical detection.