Project description:Yersinia pestis, a Gram-negative bacterium is the causative agent of the fatal communicable disease plague. The disease had a profound impact on human history. Plague bacteria are usually transmitted to humans through the bite of an infected rat flea. Earlier studies have indicated that Y. pestis can survive in environmental matrices e.g. water and soil. This study aimed to generate a peptide-based screen for identification of Y. pestis particularly from environmental matrices. We employed a shotgun proteomic approach using nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) to discover Y. pestis-specific peptides. Firstly, pure cultures of Y. pestis, Y. pseudotuberculosis, and Y. enterocolitica were grown, and their total cellular proteins were analyzed via MS, followed by in silico analysis of the obtained data. Secondly, Y. pestis-specific plasmids were analyzed using the Uniprot database, along with chromosomal-associated virulence proteins identified through extensive literature mining. To validate this screen, various concentrations of Y. pestis were spiked into the garden soil; and Y. pestis could be identified in all samples except un-spiked negative control soil sample. This study offers a valuable method for the identification of Y. pestis, by tandem mass spectrometry which may be used in environmental and clinical matrices.
