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Clin Microbiol Infect. 2017 Apr;23(4):267.e5-267.e7. doi: 10.1016/j.cmi.2016.12.009. Epub 2016 Dec 18.

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

Author information

1
Pôle de Microbiologie Médicale, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium; Service de Microbiologie, Cliniques Universitaires Saint-Luc, Brussels, Belgium. Electronic address: emmanuel.andre@uclouvain.be.
2
Pôle de Microbiologie Médicale, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
3
Service de Microbiologie, Cliniques Universitaires Saint-Luc, Brussels, Belgium.
4
Molecular and Experimental Mycobacteriology Group, Research Centre Borstel, Borstel, Germany; German Centre for Infection Research, Borstel Site, Borstel, Germany.
5
Pôle de Microbiologie Médicale, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium; Service de Microbiologie, Cliniques Universitaires Saint-Luc, Brussels, Belgium.

Abstract

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.

KEYWORDS:

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

PMID:
27998822
DOI:
10.1016/j.cmi.2016.12.009
[Indexed for MEDLINE]
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