Clinical & Research
Rationale

In 2021, one in three U.S. sepsis hospital encounters involving septic shock resulted in death.¹ In a study of 591 patients with severe sepsis or septic shock, hospital mortality was 22.9% when antibiotics were administered more than three hours after ED arrival, compared with 16.9% when administered within three hours.² In 2019, bacterial antimicrobial resistance was directly responsible for an estimated 1.27 million deaths globally.³

ISOKEM's rapid confirmation would support earlier initiation of treatment, directly addressing hospital mortality associated with delayed antibiotics, while enabling reassessment of unnecessary empirical treatment to reduce avoidable antibiotic exposure, treatment-related harm, and antibiotic resistance.

Rapid bacterial diagnostics combined with antimicrobial stewardship have been associated with $2,439 lower hospital costs per bloodstream infection and $26,298 less per surviving patient in antibiotic-resistant Gram-negative bacteremia.⁸,⁴ The latter study reported 8 fewer hospital days, while a separate health system valued each avoided inpatient day at $550 in its economic model.⁹

At the hospital level, these interventions yielded estimated annual savings of $2.34 million and $2.4 million, respectively.⁸,⁴ The broader economic burden is substantial: sepsis-related hospitalizations accounted for $52.1 billion in U.S. hospital costs and 16.7 million hospital days in 2021.¹

Earlier bacterial confirmation could reduce avoidable admissions, hospital length of stay, unnecessary investigations, and associated healthcare costs.

Conventional minimum inhibitory concentration (MIC) testing depends on successful bacterial culture and isolation, inheriting the same diagnostic delays and limitations as culture itself. In a study of 325 patients with severe manifestations of sepsis, blood-culture positivity declined from 31.4% before antibiotic administration to 19.4% afterward, a 38% relative reduction.⁵ Thus, when cultures are negative, conventional isolate-based MIC testing cannot proceed.

ISOKEM-X introduces kinetic metabolic phenotyping to measure metabolic response to antibiotic exposure directly in patient-derived specimens, without requiring prior colony formation or organism isolation. This creates a pathway toward rapid, patient-specific functional susceptibility testing within the same diagnostic window as bacterial detection, rather than waiting for sequential culture, identification, and MIC testing.

The ability to track bacterial activity in near real time opens scientifically novel and clinically useful questions. Bacterial urease expressed by urinary pathogens, for example, generates ammonia from urinary urea, raising urine pH and promoting struvite crystallization, infection stones, catheter encrustation, and urinary obstruction.⁶ Yet identifying a urease-positive organism does not establish the intensity of its pathogenic activity.

ISOKEM-X aims to characterize such enzymatic flux and its response to antibiotics to distinguish aggressive, high-activity bacterial phenotypes from less active ones. This could identify infections at greater risk of rapid crystallization and urinary obstruction, informing earlier medical intervention and escalation of care, such as urologic evaluation or catheter removal before obstruction-related complications develop.

Persistent infection may reflect ineffective antibiotic coverage, an infected device, or an undrained abscess, each requiring a different intervention.⁷

Through serial detection of viable bacteria, ISOKEM-X aims to identify persistent or recurrent infection during treatment. Earlier evidence of bacterial persistence could support timely antibiotic reassessment, device removal, abscess drainage, and other source-control interventions.

Pancreatitis, major trauma, and postoperative inflammation can produce findings that resemble infection. Fever, tachycardia, and elevated inflammatory markers, including C-reactive protein (CRP) and procalcitonin, do not establish that viable bacteria are present.

I-TRACE is being developed to add direct evidence of bacterial activity to this distinction, supporting antibiotic decisions when the clinical picture remains ambiguous.

Immunocompromised patients face a diagnostic paradox: the defenses that protect against infection also generate many of its recognizable warning signs. In chemotherapy-associated neutropenia, serious infection can progress with muted local inflammation, including little erythema or swelling.

I-TRACE targets bacterial activity rather than the strength of the immune response, with the goal of improving recognition in patients whose presentation understates their risk.

Mean arterial pressure, serum lactate, and oxygen requirements help describe how critically ill a patient is; they do not directly establish whether viable bacteria remain.

Serial I-TRACE measurements are intended to assess whether bacterial activity decreases, persists, or returns during treatment, adding evidence when clinicians must reassess antibiotic effectiveness or investigate an unresolved infection source.

An unresponsive, confused, or intubated patient may be unable to report symptoms or identify an infection source. Hypotension and altered consciousness demand immediate action but do not establish an infectious cause.

I-TRACE’s intended bedside role is to provide earlier bacteria-directed information during emergency evaluation, alongside resuscitation and empiric treatment, without waiting for culture growth.

In extraction-based LC-MS and GC-MS workflows, measuring intracellular chemistry requires stopping activity in the sampled material, extracting metabolites, and analyzing the resulting preparation. Each additional time point requires another sample and another processing cycle.

Reaction kinetics are therefore reconstructed from separately processed samples. Capturing inhibition, recovery, or a short-lived response increases both sample consumption and analytical workload. Metabolite concentration alone does not resolve this problem: the same concentration can accompany different rates of production and consumption.

KEMFlux is designed to measure compatible biochemical reactions directly and continuously through their stoichiometrically coupled products, without repeated destructive sampling of the active system.

KEMFlux’s drug-response application targets inhibition onset, magnitude, duration, and recovery within the same active system, supporting comparison of compounds and exposure schedules.

KEMFlux’s cancer-research application targets pathway responses during treatment. The objective is to distinguish functional suppression from cell loss and investigate persistent or returning activity, using reaction kinetics alongside independent viability measurements.

KEMFlux’s mitochondrial application targets selected oxidative reaction kinetics during drug exposure, oxygen limitation, and nutrient shifts, supporting investigation of changes in substrate utilization.

KEMFlux’s bioprocess application targets the onset, magnitude, and persistence of reaction responses to feeding and aeration changes, supporting feed-strategy comparison and investigation of declining process performance.