Early Detection of Acute Infections

Advanced Biomarker Identification Technology

Isomark’s scientific team continues to pursue the identification of additional disease specific biomarkers, each of which will significantly expand potential market opportunities. Isomark’s current patented technology utilizes non-invasive breath-based analysis for the early detection of acute infections that lead to septic shock. Septic shock is most frequently caused by gram negative bacteria such as E. coli or Salmonella, but in some cases infectious diseases caused by gram positive bacteria such as Staphylococcus and viruses such as Influenza create a “cytokine storm” which is detectable using Isomark technology.

Isomark’s technology identifies changes in the natural abundance of stable isotopes of carbon in expired CO₂ and changes that are detectable within 2 hours after an infecting pathogen has gained entry and initiated an immune defense in a person. As an invading pathogen infects, the body’s immune defenses are immediately mobilized. The innate immune cells, such as macrophages and neutrophils, are the first responders to bacterial invasions. When these cells encounter bacterial antigens they send out chemical messengers, such as tumor necrosis factor (TNF), Interleukin-1 beta (IL-1β), and chemokines. These messengers alert the body to the presence of infection and recruit more immune cells to fight the pathogen. As the immune system responds, resources are needed to fuel the process. TNF and IL-1β signal the catabolism of muscle tissue to free necessary amino acid (AA) resources. There are two main fates of AAs during the inflammatory response: 1) AAs can be loaded on to transfer RNA in a one step process, or 2) AAs can be used for energy or the synthesis of other immune defense compounds (e.g., nitric oxide), a process that ultimately results in the production of AA derived CO₂.

Amino acids are composed of the elements carbon, nitrogen, oxygen and sulfur. These elements exist in several isotopic states. Isomark’s technology is based on measuring the ratio of the two stable isotopic states of carbon (¹³C and ¹²C) as it is being utilized in the infectious process. In a phenomenon known as the kinetic isotope effect, heavy isotopes (¹³C) react more slowly than the lighter isotopes (¹²C). This process is called fractionation. Consider the two main fates of AAs described above. When AAs are loaded onto tRNA only one enzymatic reaction is required. When AAs are burned for energy or synthesized into other metabolites many enzymatic actions are required to convert the carbon into CO₂. Carbon produced as an end result of acute immune response is expelled as CO₂ in expired breath. Fractionation occurs during key enzymatic steps. Therefore, as the number of subsequent reactions increases the degree of fractionation also increases. The final outcome of isotopic fractionation during infectious process is a distinct change in the ¹³C/¹²C ratio that can be used as a biomarker for systemic catabolism due to infection. Since each breath represents the CO₂ produced at that moment in time, Isomark’s technology facilitates the real time monitoring of the infectious process.

The infectious process is a complex series of interrelated events. Enzymatic reactions are occurring continuously in a variety of cell types throughout the body creating a complex array of nutrient metabolites. The whole of the metabolites occurring at a single point in time is called the metabolome. During the course of an infection, the metabolome of the human body changes. New scientific data continues to be published on metabolome changes during disease states. Using these new reports, Isomark scientists are positioned to add new markers that can be detected using Isomark’s breath technology. With the addition of new markers, not only will Isomark be able to rapidly detect infection within the first few hours of the process, Isomark’s technology will provide useful diagnostic and prognosis information.