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|Title:||Target-specific assay for rapid and quantitative detection of Mycobacterium chimaera DNA|
Da Silva, A.
|Citation:||Journal of Clinical Microbiology, 2017; 55(6):1847-1856|
|Publisher:||American Society for Microbiology|
|Enrique Zozaya-Valdés, Jessica L. Porter, John Coventry, Janet A. M. Fyfe, Glen P. Carter, Anders Gonçalves da Silva, Mark B. Schultz, Torsten Seemann, Paul D. R. Johnson, Andrew J. Stewardson, Ivan Bastian, Sally A. Roberts, Benjamin P. Howden, Deborah A. Williamson, Timothy P. Stinear|
|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|
|Rights:||Copyright © 2017 American Society for Microbiology. All Rights Reserved.|
|Appears in Collections:||Biochemistry publications|
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