OBJECTIVE: To evaluate the use of denaturation high-performance liquid chromatography (dHPLC) as a rapid method to detect rifampicin (RMP) resistance based on mutations in the rpoB gene in a high-volume laboratory setting. METHODS: A total of 132 RMP-resistant Mycobacterium tuberculosis strains with different rpoB mutation were used to optimise the running condition of dHPLC as a pilot study. A blind correlation study was subsequently done between dHPLC and in vitro RMP susceptibility tests on 3167 M. tuberculosis strains in a high-throughput clinical setting. RESULTS: In the pilot study, rpoB mutation could be detected on 116/132 (87.9%) RMP-resistant strains by dHPLC. In the second phase of the study, 84/3107 (2.7%) clinical M. tuberculosis isolates were RMP-resistant. The sensitivity and specificity of dHPLC in the prediction of RMP resistance were 70/84 (83.3%) and 70/77 (91.0%), respectively. The specificity became 100% when 511 Leu to Pro mutation was excluded from the RMP resistance-related genetic changes. CONCLUSION: In the detection of RMP resistance in a high-throughput laboratory setting, dHPLC has been demonstrated to be rapid, simple, workable, automatable and inexpensive in terms of running costs and the labour involved.
OBJECTIVE: To evaluate the use of denaturation high-performance liquid chromatography (dHPLC) as a rapid method to detect rifampicin (RMP) resistance based on mutations in the rpoB gene in a high-volume laboratory setting. METHODS: A total of 132 RMP-resistant Mycobacterium tuberculosis strains with different rpoB mutation were used to optimise the running condition of dHPLC as a pilot study. A blind correlation study was subsequently done between dHPLC and in vitro RMP susceptibility tests on 3167 M. tuberculosis strains in a high-throughput clinical setting. RESULTS: In the pilot study, rpoB mutation could be detected on 116/132 (87.9%) RMP-resistant strains by dHPLC. In the second phase of the study, 84/3107 (2.7%) clinical M. tuberculosis isolates were RMP-resistant. The sensitivity and specificity of dHPLC in the prediction of RMP resistance were 70/84 (83.3%) and 70/77 (91.0%), respectively. The specificity became 100% when 511 Leu to Pro mutation was excluded from the RMP resistance-related genetic changes. CONCLUSION: In the detection of RMP resistance in a high-throughput laboratory setting, dHPLC has been demonstrated to be rapid, simple, workable, automatable and inexpensive in terms of running costs and the labour involved.
Authors: Armand Van Deun; Kya J M Aung; Valentin Bola; Rossin Lebeke; Mohamed Anwar Hossain; Willem Bram de Rijk; Leen Rigouts; Aysel Gumusboga; Gabriela Torrea; Bouke C de Jong Journal: J Clin Microbiol Date: 2013-06-12 Impact factor: 5.948
Authors: A Van Deun; L Barrera; I Bastian; L Fattorini; H Hoffmann; K M Kam; L Rigouts; S Rüsch-Gerdes; A Wright Journal: J Clin Microbiol Date: 2009-09-16 Impact factor: 5.948
Authors: Oksana Ocheretina; Lishuang Shen; Vincent E Escuyer; Marie-Marcelle Mabou; Gertrude Royal-Mardi; Sean E Collins; Jean W Pape; Daniel W Fitzgerald Journal: PLoS One Date: 2015-06-03 Impact factor: 3.240
Authors: Michel Kayomo Kaswa; Serge Bisuta; Georges Kabuya; Octavie Lunguya; André Ndongosieme; Jean Jacques Muyembe; Armand Van Deun; Marleen Boelaert Journal: PLoS One Date: 2014-04-14 Impact factor: 3.240