Background: Xpert MTB/RIF Assay is currently used in Ethiopia for the rapid diagnosis of Mycobacterium tuberculosis (MTB) and mutations that confer Rifampicin resistance. Rifampicin resistance is determined based on any mutation in the 81 bp of rpoB gene using five overlapping probes represented as Probe A (codons 507–511), Probe B (codons 512–518), Probe C (codons 518–523), Probe D (codons 523–529) and Probe E (codons 529–533). In this review, we assessed the frequency of missed probe types for Rifampicin Resistance results.
Methods: Data were reviewed from specimens received and tested using Xpert® MTB/RIF Assay at Ethiopian National Tuberculosis Reference Laboratory, in Addis Ababa from 15 July 2016 to 31 December 2018 retrospectively. All archived data were reviewed carefully to describe missed probe types and quantity of DNA in the sample.
Results: A total of 100 specimens were reported as MTB Detected Rifampicin Resistance Detected by Xpert MTB/RIF assay. More than half (55%) of these results were reported from male patients. The median age was 28.0 years (5 months to 88 years). Majorities (62%) of the cases were detected from sputum. Among the total of 38 extra pulmonary samples, lymph node aspirates were accounted 50% (19/38). The most common mutations (81.0%) were found in Probe E region followed by Probe D (10.0%), and Probe B (3.0%). Mutations in Probe A and Probe C regions were not observed. However six (6.0%) Rifampicin resistance cases were found without any missed probe type. The delta Ct max is ≥4.3. No specimen yielded Rifampicin resistance associated with more than one probe failure or mutation combinations.
Conclusion: Mutations associated with Probe E (codons 529–533) region were identified as the commonest rpoB gene mutations. The Rifampicin resistance results found without any identified missing probe needs a further study. Lower DNA amount was observed in extrapulmonary specimens compared with sputum. Keywords: Rifampicin-resistance, Molecular Beacon, DNA probes, Xpert MTB/RIF Assay