E André1, L Goeminne2, A Colmant3, P Beckert4, S Niemann4, M Delmee5. 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.
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.
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Authors: Caroline Allix-Béguec; Irena Arandjelovic; Lijun Bi; Patrick Beckert; Maryline Bonnet; Phelim Bradley; Andrea M Cabibbe; Irving Cancino-Muñoz; Mark J Caulfield; Angkana Chaiprasert; Daniela M Cirillo; David A Clifton; Iñaki Comas; Derrick W Crook; Maria R De Filippo; Han de Neeling; Roland Diel; Francis A Drobniewski; Kiatichai Faksri; Maha R Farhat; Joy Fleming; Philip Fowler; Tom A Fowler; Qian Gao; Jennifer Gardy; Deborah Gascoyne-Binzi; Ana-Luiza Gibertoni-Cruz; Ana Gil-Brusola; Tanya Golubchik; Ximena Gonzalo; Louis Grandjean; Guangxue He; Jennifer L Guthrie; Sarah Hoosdally; Martin Hunt; Zamin Iqbal; Nazir Ismail; James Johnston; Faisal M Khanzada; Chiea C Khor; Thomas A Kohl; Clare Kong; Sam Lipworth; Qingyun Liu; Gugu Maphalala; Elena Martinez; Vanessa Mathys; Matthias Merker; Paolo Miotto; Nerges Mistry; David A J Moore; Megan Murray; Stefan Niemann; Shaheed V Omar; Rick T-H Ong; Tim E A Peto; James E Posey; Therdsak Prammananan; Alexander Pym; Camilla Rodrigues; Mabel Rodrigues; Timothy Rodwell; Gian M Rossolini; Elisabeth Sánchez Padilla; Marco Schito; Xin Shen; Jay Shendure; Vitali Sintchenko; Alex Sloutsky; E Grace Smith; Matthew Snyder; Karine Soetaert; Angela M Starks; Philip Supply; Prapat Suriyapol; Sabira Tahseen; Patrick Tang; Yik-Ying Teo; Thuong N T Thuong; Guy Thwaites; Enrico Tortoli; Dick van Soolingen; A Sarah Walker; Timothy M Walker; Mark Wilcox; Daniel J Wilson; David Wyllie; Yang Yang; Hongtai Zhang; Yanlin Zhao; Baoli Zhu Journal: N Engl J Med Date: 2018-09-26 Impact factor: 91.245