Mutations in domain V of the 23S rRNA gene ofM.pneumoniae interfere with macrolide binding to rRNA, and mediate macrolide resistance.We developed a rapid and inexpensive method combining nested PCR (nPCR), single-strand conformation polymorphisms (SSCPs), and capillary electrophoresis (CE) to detect macrolide-resistant mutants.From January to December 2009 in Beijing, 665 throat swabs were collected, yielding 110 samples that tested positive for M.pneumoniae by nPCR and serological testing.We randomly selected 64 positive throat swabs for CE-SSCP analysis.The A2063G mutation was found in 57 samples, and a C2611A co-mutation was identified in one.An A2063T mutation was identified in one sample.The total mutation rate was 90.63%.All nPCR-CE-SSCP results were subjected to sequencing.The nPCR-CE-SSCP method could identify macrolide-resistant mutants directly from clinical samples.This approach would allow clinicians to rapidly choose the appropriate therapy.