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J Med Microbiol 55 (2006), 149-155; DOI: 10.1099/jmm.0.46281-0
© 2006 Society for General Microbiology
ISSN 0022-2615

Real-time detection of Mycoplasma pneumoniae in respiratory samples with an internal processing control

David Pitcher, Victoria J. Chalker, Carmen Sheppard, Robert C. George and Timothy G. Harrison

Respiratory and Systemic Infection Laboratory, Health Protection Agency Centre for Infections, 61 Colindale Avenue, London NW9 5EQ, UK

Correspondence
Victoria J. Chalker
vicki.chalker{at}hpa.org.uk

Received 4 August 2005
Accepted 16 September 2005


Real-time PCR was employed to detect a region of the P1 cytadhesin gene of Mycoplasma pneumoniae in clinical samples. An internal processing control was included that could be co-amplified simultaneously in the same reaction tube. The assay could reproducibly detect 1x103 M. pneumoniae organisms ml–1 in clinical samples. There was no amplification of DNA or signal production from 15 other species of human mycoplasmas and 19 other bacterial species. Using a panel of 175 respiratory samples taken from patients with pneumonia of proven aetiology, the sensitivity was found to be 60 % and the specificity of the assay 96·7 % when compared with serology. This assay is suitable for same-day diagnosis of M. pneumoniae infection and batch processing of respiratory samples for clinical screening.


Abbreviations: Cp, crossing point; IPC, internal processing control.

The sequence of the internal processing control bacteriophage lambda fragment is available as supplementary material in JMM Online.




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