Target-Specific Assay for Rapid and Quantitative Detection of Mycobacterium chimaera DNA

J Clin Microbiol. 2017 Jun;55(6):1847-1856. doi: 10.1128/JCM.00197-17. Epub 2017 Apr 5.

Abstract

Mycobacterium chimaera is an opportunistic environmental mycobacterium belonging to the Mycobacterium avium-M. intracellulare complex. Although most commonly associated with pulmonary disease, there has been growing awareness of invasive M. chimaera infections following cardiac surgery. Investigations suggest worldwide spread of a specific M. chimaera clone, associated with contaminated hospital heater-cooler units used during the surgery. Given the global dissemination of this clone, its potential to cause invasive disease, and the laboriousness of current culture-based diagnostic methods, there is a pressing need to develop rapid and accurate diagnostic assays specific for M. chimaera Here, we assessed 354 mycobacterial genome sequences and confirmed that M. chimaera is a phylogenetically coherent group. In silico comparisons indicated six DNA regions present only in M. chimaera We targeted one of these regions and developed a TaqMan quantitative PCR (qPCR) assay for M. chimaera with a detection limit of 100 CFU/ml in whole blood spiked with bacteria. In vitro screening against DNA extracted from 40 other mycobacterial species and 22 bacterial species from 21 diverse genera confirmed the in silico-predicted specificity for M. chimaera Screening 33 water samples from heater-cooler units with this assay highlighted the increased sensitivity of PCR compared to culture, with 15 of 23 culture-negative samples positive by M. chimaera qPCR. We have thus developed a robust molecular assay that can be readily and rapidly deployed to screen clinical and environmental specimens for M. chimaera.

Keywords: diagnostics; genomics; infectious disease; mycobacterium.

Publication types

  • Evaluation Study

MeSH terms

  • DNA, Bacterial / genetics*
  • DNA, Bacterial / isolation & purification*
  • Humans
  • Molecular Diagnostic Techniques / methods*
  • Mycobacterium / genetics*
  • Mycobacterium / isolation & purification*
  • Mycobacterium Infections / diagnosis*
  • Mycobacterium Infections / microbiology
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • DNA, Bacterial