Determining Viability of M. ulcerans by 16S rRNA RT Reverse Transcriptase Real-Time PCR

To overcome drawbacks of M. ulcerans culture in terms of incubation time and low sensitivity for the detection of viable bacilli from clinical specimens, a highly sensitive and M. ulcerans-specific RNA-based viability assay was developed. The assay combines a 16S rRNA reverse transcriptase real-time PCR (16S rRNA RT qPCR) to determine bacterial viability with an IS2404 quantitative real-time PCR (IS2404 qPCR) to ensure specificity as well as simultaneous quantification of bacilli. This proved to be highly efficient in detecting viable bacilli in clinical samples when implemented in the field.