Novel rapid PCR for the detection of Ile491Phe rpoB mutation of Mycobacterium tuberculosis, a rifampicin-resistance-conferring mutation undetected by commercial assays

作者：Andre E^*; Goeminne L; Colmant A; Beckert P; Niemann S; Delmee M

来源：Clinical Microbiology and Infection, 2017, 23(4): 267E5-267E7.

DOI：10.1016/j.cmi.2016.12.009

• I491F
• I572F
• Ile491Phe
• Ile572Phe
• Multidrug-resistant tuberculosis
• Multiplex allele-specific-PCR
• rpoB
• Tuberculosis
• Xpert

Objectives: Neither the liquid medium-based Bactec MGIT, nor commercial molecular assays such as the Xpert MTB/RIF and the MTBDRplus V2.0 assays are capable of detecting up to 30% of rifampicin-resistant Mycobacterium tuberculosis strains in Swaziland because of the large proportion of the rpoB Ile491Phe mutations. In other countries, the frequency of this mutation is thought to be low.
Methods: We designed a real-time multiplex allele-specific PCR assay to identify the rpoB Ile491Phe mutation responsible for these undetected resistant M. tuberculosis strains.
Results: The technique showed 100% similarity with rpoB sequencing on a panel of 78 strains from Swaziland.
Conclusions: We propose that the detection of the rpoB Ile491Phe rpoB mutation should complement commercial assays for the diagnosis of rifampicin-resistant M. tuberculosis in routine conditions, particularly in countries where this specific mutation is frequent. The technique proposed in this paper is adapted for most reference laboratories.

• 出版日期2017-4