MEASURE: Multiplex Exhaled Breath Condensate - Scanning Using Rapid Electro-Analytics.

Exhaled breath condensate is an emerging source of inflammatory biomarkers suitable for the noninvasive detection of respiratory disorders. Current gold standard methods are highly invasive and pose challenges in sample collection during airway inflammation monitoring. Cytokine biomarkers are detectable in EBC at increased or decreased concentrations. IL-6, IL-1ß, IL-8, and hs-CRP are characteristic biomarkers identified in respiratory disorders. We have demonstrated the promising outcomes of a 16-plexed electrochemical platform - READ 2.0 for the multiplexed detection of characteristic biomarkers in EBC. The sensor demonstrates dynamic ranges of 1-243 pg/mL with a lower detection limit of 1 pg/mL for IL-6 and IL-1ß, while the detection range and limit of detection for IL-8 and hs-CRP is 1-150 pg/mL and 3 pg/mL, respectively. The detection accuracies for the biomarkers are in the range of ∼85 ± 15% to ∼100 ± 10%. The sensor shows a nonspecific response to similar cross-reacting biomarkers. Analytical validation of the sensor with ELISA as the standard reference generated a correlation of R2 > 0.96 and mean biases of 10.9, 3.5, 17.4, and 3.9 pg/mL between the two methods for IL-6, IL-1ß, IL-8, and hs-CRP, respectively. The precision of the sensor in detecting low biomarker concentrations yields a %CV of <7%. The variation in the sensor's response on repeat EBC sample measurements and within a 6 h duration is less than 10%. The READ 2.0 platform shows a promise that EBC-based biomarker detection can prove to be vital in predicting the severity and survival rates of respiratory disorders and serve as a reference point for monitoring EBC-based biomarkers.

EBC-SURE (exhaled breath condensate- scanning using rapid electro analytics): A non-faradaic and non-invasive electrochemical assay to screen for pro-inflammatory biomarkers in human breath condensate.

The innovation of this work lies in the trace detection of inflammatory biomarkers (IL-6, hs-CRP) in human exhaled breath condensate on the developed EBC-SURE platform as a point-of-care aid for respiratory disorder diagnosis. The unique design of the EBC-SURE leverages non-faradaic electrochemical impedance spectroscopy to capture target-specific biomolecular interactions for highly sensitive biomarker detection. For sensor calibration, EBC-SURE's performance is assessed to measure the response of the sensor to a known concentration by spike and recovery analysis with a recovery error of <20% and an extended dynamic range over 3-log orders. The lowest detection limits for IL-6 and hs-CRP detection in EBC were found to be 3.2 pg/mL and 4 pg/mL respectively. The intra-assay and inter-assay efficacy of EBC-SURE for its usage as a diagnostic device was established through repeatability and reproducibility (over 48 h s) performance testing. The percentage variations (<20%) met the Clinical and Laboratory Standards Institute standards (CLSI) indicating a highly stable performance for robust biomarker detection. EBC-SURE generated highly selective IL-6 and hs-CRP responses in the presence of other non-specific cytokines. Statistical validation methods- Correlation and Bland Altman analysis established the one-to-one agreement between EBC-SURE and the reference method. Correlation analysis generated a Pearson's R value of 0.99 for IL-6 and hs-CRP. Bland-Altman analysis indicated a good agreement between both the methods with all data points confined within the ±2SD limits. We have demonstrated EBC-SURE's ability in detecting inflammatory biomarkers in human breath condensate towards developing a non-invasive technology that can quantify biomarker levels associated with healthy and acute inflammatory conditions.

Point-of-use sweat biosensor to track the endocrine-inflammation relationship for chronic disease monitoring.

AIM: The hypothalamic-pituitary-adrenal axis is involved in maintaining homeostasis by engaging with the parasympathetic nervous system. During the process of disease affliction, this relationship is disturbed and there is an imbalance driven response observed. MATERIALS & METHODS: By monitoring the two key components involved in these pathways, cortisol and TNF-α, the manifestations of chronic stress on the body's homeostasis can be evaluated in a comprehensive manner. This work highlights the development of an electrochemical detection system for the two biomarkers through human sweat. RESULTS: Limit of detection and dynamic ranges are 1 ng/ml, 1-200 ng/ml for cortisol and 1 pg/ml, 1-1000 pg/ml for TNF-α. CONCLUSION: This wearable system is designed to be a point of use, chronic disease self-monitoring and management platform.