Rapid Detection of multiple sepsis pathogens

Abstract Sepsis is a major public health concern and the primary cause of death in the intensive care unit accounting for more than $ billion of total US hospital costs in Sepsis can lead to a life threatening drop in blood pressure termed septic shock which can quickly lead to the failure of vital organs such as lungs kidneys and liver and eventually to death Nearly all sepsis cases are associated with bacterial or fungal blood stream infection BSI Due to the rapid progression of sepsis the therapeutic window is very short In order for hospitals to qualify for Medicare in the US physicians are required to treat suspected BSI within hours of an empirical BSI diagnosis The current standard of care until definite diagnosis of pathogen is reached by immediate intervention with broad spectrum empirical antimicrobial therapy and supportive treatment have been shown to have the greatest impact on outcomes However the broad spectrum drugs are not always well tolerated often do not target fungal infections and are expensive Therefore a major hurdle of treating sepsis caused by BSI remains reaching a fast diagnosis of the causative pathogen A variety of nucleic acid based molecular tests can be performed on positive cultures for identifying the pathogen s and characterizing their antibiotic susceptibility However the average time to receive a positive culture result is slow days and species identification and drug sensitivity profiling prolongs the process even further We offer the use of the TangenDx Molecular Diagnostic Platform for BSI Detection Tangen s unique technological advantage is in capturing and concentrating the pathogens from a raw sample before they go into the instrument Tangen is able to achieve high sensitivity of detection directly from whole blood because of development of unique laser drilled kapton filters that enable large blood volumes ranging from tens of milliliters to tens of microliters to be concentrated by the LVCTM technology For BSI the TangenDx will provide a diagnostic sensitivity that is equal to or superior to blood culture methods and deliver a diagnosis and pathogen ID in less than hours total A major hurdle to using nucleic acid based molecular assays to directly detect blood stream pathogens is the fact that the concentration of cells per ml of blood is very low CFU ml of blood Our approach to overcome this problem involves two strategies first targeting multi copy gene regions e g Mitochondrial transposons or rRNA genes and second performing a global Whole Genome Amplification step in the central chamber of the Assay disk prior to distributing the amplified genomic material to the Assay wells We have demonstrated that it is feasible to detect as few as genome copies with LAMP primers targeting rRNA multi copy genes and that WGA can be successfully applied to the captured genomes prior to the LAMP reactions We will also test the fully integrated system with spiked blood samples Such a rapid diagnostics with faster turnaround time could enable rapid selection of appropriate treatment and reduce the to mortality sepsis rate Narrative Sepsis occurs when an infection induces inflammatory responses throughout the body and in severe cases one or more organs fail It is a major public health concern and the primary cause of death in the intensive care unit Nearly all sepsis cases are associated with bacterial or fungal blood stream infection Due to the rapid progression of sepsis the therapeutic window is very short The TangenDx platform enables the quick analysis of blood samples and the detection of a panel of pathogens present in the blood within two hour Such a rapid and highly sensitive test for BSI has the potential to achieve lower mortality and better patient outcomes