Target-Specific Assay for Rapid and Quantitative Detection of Mycobacterium chimaera DNA
| Autor: | Jessica L. Porter, Andrew J. Stewardson, Janet A. M. Fyfe, Paul D R Johnson, John Coventry, Glen P. Carter, Enrique Zozaya-Valdés, Sally Roberts, Torsten Seemann, Mark B. Schultz, Anders Gonçalves da Silva, Timothy P. Stinear, Deborah A Williamson, Benjamin P Howden, Ivan Bastian |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Microbiology (medical) DNA Bacterial Mycobacterium Infections biology In silico Genomics Mycobacteriology and Aerobic Actinomycetes biology.organism_classification Genome Polymerase Chain Reaction Sensitivity and Specificity law.invention Microbiology Mycobacterium 03 medical and health sciences 030104 developmental biology Real-time polymerase chain reaction Molecular Diagnostic Techniques law TaqMan Humans Polymerase chain reaction Bacteria |
| Zdroj: | Journal of clinical microbiology. 55(6) |
| ISSN: | 1098-660X |
| Popis: | 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 . |
| Databáze: | OpenAIRE |
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