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Rapid pathogen diagnostic through amplification of red blood cell-associated DNA
Sepsis, an overwhelming and life-threatening response to infection, is the most common cause of death in US hospitals. Sepsis is associated with a mortality rate of 25–30% and mortality due to septic shock is 50–85%. Rapid diagnosis and intervention in sepsis are essential given an increased mortality rate of 8% for every hour of delayed treatment. Current therapy involves early administration of broad-spectrum antibiotics to neutralize sepsis-causing bacteria and initiation of bacterial cultures to inform more targeted downstream treatment. These cultures can often take days to return results, and patients would benefit from a faster diagnostic tool.
Although direct bacterial genome amplification from blood has been proposed as a way to identify pathogens, previous attempts from patient plasma or serum have failed. This technology provides a method to enrich for bacterial or viral DNA, enabling pathogen identification from a single drop of patient blood in less time than is required for standard cultures.
Recently, the Mangalmurti lab discovered that red blood cells (RBCs) bind cell free CpG-containing mitochondrial DNA. They have now shown that RBCs also bind CpG DNA from invading bacteria or viruses, and have harnessed this phenomenon to detect pathogens in the blood. In this method, DNA is isolated from purified RBCs and amplified using pathogen-specific primers against 16s rRNA gene DNA. Positive signals identify the invading pathogen and are used to inform targeted therapy. This entire process can be completed within a few hours, and simple substitution of primers for alternative targets support broad detection capabilities.
Diagnosis up to two days faster than standard blood cultures
Faster termination of broad-spectrum antibiotic treatment will reduce likelihood of antibiotic resistance and emergence of multi-drug resistant pathogens
Direct sequencing can detect non-bacterial pathogens including viruses and parasites
Procedures within the method can be performed by clinical microbiology or clinical pathology labs
Stage of Development:
Proof of Concept
Hotz, MJ et al.;
Am J Respir Crit Care Med 2018 Feb 15; 197(4): 470.
Anderson, et al.
J Immunol, 2018, 201(5)-1343.
Docket # 18-8596
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