Changes in prevalence and load of airway bacteria using quantitative PCR in stable and exacerbated COPD
- Davinder S Garcha1,
- Sarah J Thurston1,
- Anant R C Patel1,
- Alexander J Mackay1,
- James J P Goldring1,
- Gavin C Donaldson1,
- Timothy D McHugh2,
- Jadwiga A Wedzicha1
- 1Centre for Respiratory Medicine, University College London, London, UK
- 2Centre for Clinical Microbiology, University College London, London, UK
- Correspondence to Davinder S Garcha, Centre for Respiratory Medicine, University College London, London NW3 2PF, UK;
Contributors Study design: DSG, GCD, TDM, JAW; sputum collection and processing: ARCP, AJM, JJPG; data analysis: DSG, GCD; DNA extraction: DSG, SJT; PCR: DSG; manuscript drafting & revision: DSG, ARCP, AJM, GCD, TDM, JAW; funding: JAW.
- Received 18 March 2012
- Accepted 10 July 2012
- Published Online First 3 August 2012
Background Prevalence and load of airway bacteria in stable and exacerbated chronic obstructive pulmonary disease (COPD) has been previously studied using microbiological culture. Molecular techniques, such as quantitative PCR (qPCR), may be more informative.
Methods In this study, 373 sputum samples from 134 COPD outpatients were assessed for prevalence and load of typical airway bacteria (Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis) by multiplex qPCR, with 176 samples analysed for atypical bacteria. Paired stable and exacerbation typical bacteria data were compared in 52 patients. We compared routine culture with qPCR in 177/373 samples.
Results Typical bacteria were more prevalent in exacerbation than stable-state paired samples: 30/52 (57.7%) vs. 14/52 (26.9%); p=0.001. In patients who were bacteria-positive at both time points, mean (±1 SEM) load was significantly higher at exacerbation than stable state (108.5(±0.3) vs. 107.2(±0.5) cfu/ml), constituting a 20-fold increase (p=0.011). qPCR was more discriminatory at detecting typical bacteria than microbiological culture (prevalence 59.3% vs. 24.3%; p<0.001). At stable state, higher airway bacterial load correlated with more severe airflow limitation (FEV1%predicted) (r=−0.299; p=0.033) and higher inhaled corticosteroid dosage (r=0.382; p=0.008). Mean C-reactive protein was higher in bacterial-associated exacerbations (35.0 Vs 25.1 mg/L; p=0.032).
Conclusions Airway bacterial prevalence and load increase at COPD exacerbations and are an aetiological factor. qPCR is more discriminatory than culture, identifying higher airway bacterial prevalence. Exacerbations associated with bacterial detection showed a higher mean C-reactive protein level. In the stable state, airway bacterial load is related to more severe airflow limitation and higher inhaled corticosteroid dosage used.
Funding This work was supported by Medical Research Council (UK).
Competing interests None.
Ethics approval Ethics approval was provide by Royal Free Research Ethics Committee.
Provenance and peer review Not commissioned; externally peer reviewed.