Clair L Preece1, Audrey Perry2, Bethany Gray2, Dervla T Kenna3, Amanda L Jones4, Stephen P Cummings4, Ali Robb2, Matthew F Thomas5, Malcolm Brodlie5, Christopher J O'Brien5, Stephen J Bourke6, John D Perry7. 1. Microbiology Department, Freeman Hospital, Newcastle upon Tyne NE7 7DN, UK; Faculty of Health and Life Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK. 2. Microbiology Department, Freeman Hospital, Newcastle upon Tyne NE7 7DN, UK. 3. Antimicrobial Resistance and Healthcare Associated Infections Reference Unit, Public Health England, London, UK. 4. Faculty of Health and Life Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK. 5. Paediatric Respiratory Unit, Great North Children's Hospital, Newcastle upon Tyne NE1 4LP, UK. 6. Adult Cystic Fibrosis Centre, Royal Victoria Infirmary, Newcastle upon Tyne NE1 4LP, UK. 7. Microbiology Department, Freeman Hospital, Newcastle upon Tyne NE7 7DN, UK; Faculty of Health and Life Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK. Electronic address: john.perry@nuth.nhs.uk.
Abstract
BACKGROUND: Isolation of mycobacteria from the sputum of patients with cystic fibrosis (CF) is challenging due to the overgrowth of cultures by other bacteria and fungi. In this setting, Burkholderia cepacia selective agar (BCSA) has been recommended as a convenient and effective culture medium for the isolation of rapidly-growing, non-tuberculous mycobacteria (NTM). A novel selective culture medium (RGM medium) was evaluated for the isolation of rapidly-growing NTM from the sputum of children and adults with CF. METHODS: A total of 118 isolates of rapidly-growing mycobacteria and 98 other bacteria and fungi were inoculated onto RGM medium. These were assessed for growth at 30°C over a seven day period. A total of 502 consecutive sputum samples were collected from 210 patients with CF. Each sample was homogenized and cultured onto RGM medium and also onto BCSA. Cultures were incubated for 10days at 30°C. RESULTS: Of 118 isolates of mycobacteria all but one grew well on RGM medium, whereas 94% of other bacteria and fungi were inhibited. A total of 55 sputum samples (from 33 distinct patients) yielded NTM using a combination of both RGM and BCSA (prevalence: 15.7%). NTM were recovered from 54 sputum samples using RGM medium compared with only 17 samples using BCSA (sensitivity 98% vs. 31%; P≤0.0001). A total of 419 isolates of non-mycobacteria were recovered from sputum samples on BCSA compared with 46 on RGM medium. CONCLUSIONS: RGM medium offers a simple and effective culture method for the isolation of rapidly-growing mycobacteria from sputum samples from patients with CF without decontamination of samples. RGM medium allows for the systematic screening of all sputum samples routinely referred for culture from patients with CF.
BACKGROUND: Isolation of mycobacteria from the sputum of patients with cystic fibrosis (CF) is challenging due to the overgrowth of cultures by other bacteria and fungi. In this setting, Burkholderia cepacia selective agar (BCSA) has been recommended as a convenient and effective culture medium for the isolation of rapidly-growing, non-tuberculous mycobacteria (NTM). A novel selective culture medium (RGM medium) was evaluated for the isolation of rapidly-growing NTM from the sputum of children and adults with CF. METHODS: A total of 118 isolates of rapidly-growing mycobacteria and 98 other bacteria and fungi were inoculated onto RGM medium. These were assessed for growth at 30°C over a seven day period. A total of 502 consecutive sputum samples were collected from 210 patients with CF. Each sample was homogenized and cultured onto RGM medium and also onto BCSA. Cultures were incubated for 10days at 30°C. RESULTS: Of 118 isolates of mycobacteria all but one grew well on RGM medium, whereas 94% of other bacteria and fungi were inhibited. A total of 55 sputum samples (from 33 distinct patients) yielded NTM using a combination of both RGM and BCSA (prevalence: 15.7%). NTM were recovered from 54 sputum samples using RGM medium compared with only 17 samples using BCSA (sensitivity 98% vs. 31%; P≤0.0001). A total of 419 isolates of non-mycobacteria were recovered from sputum samples on BCSA compared with 46 on RGM medium. CONCLUSIONS: RGM medium offers a simple and effective culture method for the isolation of rapidly-growing mycobacteria from sputum samples from patients with CF without decontamination of samples. RGM medium allows for the systematic screening of all sputum samples routinely referred for culture from patients with CF.
Authors: Suwatchareeporn Rotcheewaphan; Oluwadamilola E Odusanya; Christina M Henderson; Dominic Stephenson; Kenneth N Olivier; John D Perry; Adrian M Zelazny Journal: J Clin Microbiol Date: 2019-01-30 Impact factor: 5.948
Authors: Clair L Preece; Thomas A Wichelhaus; Audrey Perry; Amanda L Jones; Stephen P Cummings; John D Perry; Michael Hogardt Journal: J Clin Microbiol Date: 2016-04-20 Impact factor: 5.948
Authors: Betty A Forbes; Geraldine S Hall; Melissa B Miller; Susan M Novak; Marie-Claire Rowlinson; Max Salfinger; Akos Somoskövi; David M Warshauer; Michael L Wilson Journal: Clin Microbiol Rev Date: 2018-01-31 Impact factor: 26.132
Authors: Aaron I Gardner; Elliot McClenaghan; Gemma Saint; Paul S McNamara; Malcolm Brodlie; Matthew F Thomas Journal: Clin Infect Dis Date: 2019-02-15 Impact factor: 9.079
Authors: Anaïs Scohy; Sophie Gohy; Vanessa Mathys; Guillaume Sapriel; Laëtitia Toussaint; Florian Bressant; Ali Zitouni; Marie-Noël Teylaert; Marie-Christine Vander Meeren; Alexandre Colmant; Anne Simon; John D Perry; Patrick Lebecque; Emmanuel André Journal: J Clin Tuberc Other Mycobact Dis Date: 2018-08-01
Authors: H Al-Momani; A Perry; R Jones; S Bourke; S Doe; J Perry; A Anderson; T Forrest; I Forrest; M Griffin; M Brodlie; J Pearson; C Ward Journal: Sci Rep Date: 2017-04-24 Impact factor: 4.379