Label- and amplification-free electrochemical detection of bacterial ribosomal RNA

Current approaches to molecular diagnostics rely heavily on PCR amplification and optical detection methods which have restrictions when applied to point of care (POC) applications. Herein we describe the development of a label-free and amplification-free method of pathogen detection applied to Escherichia coli which overcomes the bottleneck of complex sample preparation and has the potential to be implemented as a rapid, cost effective test suitable for point of care use. Ribosomal RNA is naturally amplified in bacterial cells, which makes it a promising target for sensitive detection without the necessity for prior in vitro amplification. Using fluorescent microarray methods with rRNA targets from a range of pathogens, an optimal probe was selected from a pool of probe candidates identified in silico. The specificity of probes was investigated on DNA microarray using fluorescently labeled 16S rRNA target. The probe yielding highest specificity performance was evaluated in terms of sensitivity and a LOD of 20 pM was achieved on fluorescent glass microarray. This probe was transferred to an EIS end point format and specificity which correlated to microarray data was demonstrated. Excellent sensitivity was facilitated by the use of uncharged PNA probes and large 16S rRNA target and investigations resulted in an LOD of 50 pM. An alternative kinetic EIS assay format was demonstrated with which rRNA could be detected in a species specific manner within 10-40 min at room temperature without wash steps

Use of an innovative personality-mindset profiling tool to guide culture-change strategies among different healthcare worker groups

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Derived personality profiles and messaging strategies for each HCW clinical-contact category based on the ColourGrid^® profiles shown in Fig 3.

<p>Derived personality profiles and messaging strategies for each HCW clinical-contact category based on the ColourGrid^® profiles shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0140509#pone.0140509.g003" target="_blank">Fig 3</a>.</p

ColourGrid^® profiles for each of the three doctor categories based on PS data.

<p>The number of HCWs analysed in each group are shown in the lower right-hand corner of each matrix. Where box sizes >150%, the percentage is stated. Derived personality profiles and messaging strategies are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0140509#pone.0140509.t004" target="_blank">Table 4</a>.</p

ColourGrid^® principles and summary profiles.

<p>The principles of ColourGrid are based on Hofstede’s cultural dimensions theory (see text). The principles include: <b>Power distance:</b> relates to the extent to which the less powerful members of organizations and institutions accept and expect that power is distributed unequally. It suggests that a society’s level of inequality is endorsed by the followers as much as by the leaders. <b>Uncertainty avoidance:</b> indicates to what extent society tolerates uncertainty and ambiguity, and it shows how comfortable its members feel in unstructured situations which are novel, unknown, surprising or different from usual. <b>Individualism:</b> is the degree to which individuals are integrated into tight groups (collectivist) or loose groups (individualist). <b>Long-term orientation:</b> reflects long-term pragmatic attitudes versus short-term normative attitudes. Cultures scoring high on this dimension show emphasis on future rewards, notably saving, persistence, and adapting to changing circumstances.</p