A Loop-mediated Isothermal Amplification Method for Rapid Direct Detection and Differentiation of Non-pathogenic and Verocytotoxigenic Escherichia coli in Beef and Bovine Faeces

The aim of this study was to develop a multiplex loop-mediated isothermal amplification (LAMP) method capable of detecting Escherichia coli generally and verocytotoxigenic E. coli (VTEC) specifically in beef and bovine faeces. The LAMP assay developed was highly specific (100%) and able to distinguish between E. coli and VTEC based on the amplification of the phoA, and stx1 and/or stx2 genes, respectively. In the absence of an enrichment step, the limit of detection 50% (LOD50) of the LAMP assay was determined to be 2.83, 3.17 and 2.83-3.17 log CFU/g for E. coli with phoA, stx1 and stx2 genes, respectively, when artificially inoculated minced beef and bovine faeces were tested. The LAMP calibration curves generated with pure cultures, and spiked beef and faeces, suggested that the assay had good quantification capability. Validation of the assay, performed using retail beef and bovine faeces samples, demonstrated good correlation between counts obtained by the LAMP assay and by a conventional culture method, but suggested the possibility of false negative LAMP results for 12.5-14.7% of samples tested. The multiplex LAMP assay developed potentially represents a rapid alternative to culture for monitoring E.coli levels in beef or faeces and it would provide additional information on the presence of VTEC. However, some further optimisation is needed to improve detection sensitivity.

Conclusion: patterns of non-territorial autonomy

This article seeks to generalise about the significance of non-territorial autonomy as a mechanism for the management of ethnic conflict on the basis of a set of case studies covering the Ottoman empire and its successor states, the Habsburg monarchy, the Jewish minorities of Europe, interwar Estonia, contemporary Belgium, and two indigenous peoples, the Sami in Norway and Maori in New Zealand. It begins by assessing the extent to which the spatial distribution of ethnonational communities determined the range of autonomy options available—whether these might be territorial or whether only non-territorial autonomy would be realistic. The article continues with an assessment of the significance of ‘autonomy’ in circumstances where the institutions with which it is associated enjoy a non-territorial rather than a territorial writ. It concludes by suggesting that in almost all cases where autonomy is extended to a minority within a state this is exercised on a territorial basis, and that in many cases of non-territorial autonomy, or national–cultural autonomy, the powers assumed by the ‘autonomous’ institutions are substantially symbolic. It argues that notwithstanding the limited empirical evidence for the existence of non-territorial autonomy, this device should not be written off at a normative level.