MPLEx: a method for simultaneous pathogen inactivation and extraction of samples for multi-omics profiling.

摘要: The continued emergence and spread of infectious agents is great concern, systems biology approaches to disease research can advance our understanding host-pathogen relationships facilitate the development new therapies vaccines. Molecular characterization samples outside appropriate biosafety containment take place only subsequent pathogen inactivation. Herein, we describe a modified Folch extraction using chloroform/methanol that facilitates molecular by enabling simultaneous inactivation proteins, metabolites, lipids for mass spectrometry-based multi-omics measurements. This single-sample metabolite, protein lipid (MPLEx) method resulted in complete clinically important bacterial viral pathogens with exposed membranes, including Yersinia pestis, Salmonella Typhimurium, Campylobacter jejuni pure culture, jejuni, West Nile, MERS-CoV, Ebola, influenza H7N9 viruses infection studies. In addition, >99% inactivation, which increased solvent exposure time, was also observed without membranes community-associated methicillin-resistant Staphylococcus aureus, Clostridium difficile spores vegetative cells, adenovirus type 5. overall pipeline proteomic, metabolomic, lipidomic analyses evaluated human epithelial lung cell line infected wild-type mutant viruses, thereby demonstrating MPLEx yields biomaterial sufficient quality analyses. Based on these experimental results, believe will studies measurements from single specimen high success membranes.