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Doubling times were calculated as described by Khalichi et al. ATP hydrolysis activity assay. Reactions were stopped at time points 0, 5 and 10 min by removing an aliquot to iced stop buffer [1. Assays were performed in quadruplicate.
GAPDH assay of whole cells. Cells were harvested by centrifugation and suspended to OD600 1. Cells were removed by centrifugation and A340 of the supernatants was read and standardized by dividing this value by the measured OD600. Assays were performed in triplicate for each strain.
Comparison of cell-surface-localized P1 by whole cell dot-blot. Wild-type and YidC mutant strains were compared for their reactivity with a panel of extensively characterized monoclonal antibodies McArthur et al. Evaluation of S. Adherence of S. For evaluation of growth on agar, cells were grown 16 h in TDM-cellobiose and diluted 1 : 10 into fresh TDM-cellobiose and grown to mid-exponential phase. They were then streaked on TDM agar containing 0.
Western blot analysis. Cells were lysed with glass beads 0. Evaluation of bacterial growth. Cultures of S.
Doubling times were calculated as described by Khalichi et al. ATP hydrolysis activity assay.
Reactions were stopped at time points 0, 5 and 10 min by removing an aliquot to iced stop buffer [1. Assays were performed in quadruplicate.
GAPDH assay of whole cells. Cells were harvested by centrifugation and suspended to OD600 1.
This was identical to model 3, but patient data was collected over a 3 month period. Model 4 clusters and sizes were similar to those found in the previous word using model 3. After applying the definitions as described in Table 2 , only 1 patient was possibly related to another patient.
All other patients were impossible to relate in time and place Table 6.
However, despite this extra level of clustering, epidemiological relationships between these isolates and patients could still not be identified. In outbreak settings, newer typing techniques such as whole genome sequencing WGS are proving to be helpful in healthcare- related transmission events settings, and are able to distinguish outbreak from non-outbreak bacterial strains [ 21 , 22 ].
However, this technique still needs threshold analyses for defining recent transmissions. Also, currently, the WGS technique is not generally available for use in routine patient settings due to the fact that it is a complex, laborious, time-consuming and expensive technique.
In this publication, clinical and molecular epidemiology both genetic and phenotypic data have been combined in an attempt to detect healthcare-related transmission events in a non-outbreak setting. In the Netherlands, recently introduced guidelines for multidrug-resistant microorganisms stated that all ESBL-producing E.
Additionally, it is not clear how any results obtained should actually be interpreted. In a systematic review, Kramer et al.