Development and evaluation of a detection method for 23S rRNA mutation of Bordetella pertussis with allele-specific PCR

  Objective  To establish a highly specific and sensitive method to detect 23S rRNA mutation in Bordetella pertussis (B. pertussis) with allele-specific PCR (AS-PCR).

  Methods  Based on the proven association of erythromycin resistance with the A2047G mutation in the 23S rRNA of B. pertussis, primers containing specific mutation sites were designed to establish the two-stage PCR. High performance capillary electrophoresis was employed to detect the target band to identify the mutation of 23S rRNA.

  Results  Assessed by the golden standard e.g. the results of gene sequencing, the sensitivity is 96% (144/150) and the specificity is 100% (100/100), with a kappa value of 95 (P < 0.01) for the detection of the mutant strains using the established AS-PCR method; while, the sensitivity is 100% (18/18) and the specificity is 100% (100/100), with a kappa value of 1 (P < 0.01) for the detection of wild strains.

  Conclusion  The established AS-PCR method is highly sensitive and specific for the detection of 23S rRNA A2047G mutation in Bordetella pertussis; the method could be applied to rapid detection of rythromycin-resistance of Bordetella pertussis in common clinical microbiology laboratories.