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Volume 19, Number 7—July 2013

Dispatch

Undetected Multidrug-Resistant Tuberculosis Amplified by First-line Therapy in Mixed Infection

Suzanne M. Hingley-Wilson1Comments to Author , Rosalyn Casey1, David Connell, Samuel Bremang, Jason T. Evans, Peter M. Hawkey, Grace E. Smith, Annette Jepson, Stuart Philip, Onn Min Kon, and Ajit Lalvani
Author affiliations: Imperial College London, London, UK (S.M. Hingley-Wilson, R. Casey, D. Connell, S. Bremang, A. Lalvani); Heart of England National Health Service Foundation Trust, Birmingham, UK (J.T. Evans, P.M. Hawkey, G.E. Smith); School of Infection and Immunity, University of Birmingham, Birmingham (P.M. Hawkey); Imperial College National Health Service HealthcareTrust, London (A. Jepson, S. Philip, O.M. Kon); 1Current affiliation: University of Surrey, Guildford, UK.

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Figure 2

Analyses of the amplification of multidrug-resistant Mycobacterium tuberculosis during treatment of a drug-sensitive (S) strain in a mixed infection (i.e., infection with drug-resistant [R] and S strains). In the presence of isoniazid (INH), the faster growing S strain lost its competitive advantage, and the R strain became more prevalent. A–C) Data are means of 3 independent replicates with SE bars. A) Single strain growth analyses of S (black circles) and R (gray triangles) M. tuberculosis str

Figure 2. . Analyses of the amplification of multidrug-resistant Mycobacterium tuberculosis during treatment of a drug-sensitive (S) strain in a mixed infection (i.e., infection with drug-resistant [R] and S strains). In the presence of isoniazid (INH), the faster growing S strain lost its competitive advantage, and the R strain became more prevalent. A–C) Data are means of 3 independent replicates with SE bars. A) Single strain growth analyses of S (black circles) and R (gray triangles) M. tuberculosis strains. Growth was measured by optical density at 600 nm (OD600). *p<0.05. B) Competitive growth analyses of mixed strains alone (black squares) and with 0.2 μg/mL INH (gray triangles). Growth, in triplicate, in 7H9 broth plus OADC (oleic acid, albumin, dextrose, and catalase growth supplement), glycerol, and Tween 80 was measured by optical density at OD600. Statistical analyses were performed on triplicate samples by using 2-way analysis of variance. *p<0.05. C) Identification of the predominant strain in mixed cultures with and without 0.2 μg/mL INH (INH/plate+ and INH/plate−, respectively). Strains were identified on day 7 by plating a 10-fold dilution series of co-cultures onto 7H10 agar, plus OADC and glycerol, with or without 0.2 μg/mL INH (0/plate+ and 0/plate−, respectively) and incubating for 2 weeks at 37°C. Statistical analyses were conducted on triplicate samples by using a 2-tailed t-test. *p<0.02. CFU, colony-forming units.

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