MiRNA analysis simplified using multi-ChemiRNA Tech.

Ultra-precise quantification of DILI-associated circulating miR-122, providing an accurate assessment of liver injury.

Drug-induced liver injury (DILI) is a potentially fatal adverse event and a leading cause for pre- and post-marketing drug withdrawal. Several multinational DILI initiatives have now recommended a panel of protein and microRNA (miRNA) biomarkers that can detect early liver injury and inform about mechanistic basis. This manuscript describes the development of seqCOMBO, a unique combo-multiplexed assay which combines the dynamic chemical labelling approach and an antibody-dependant method on the Luminex MAGPIX system. SeqCOMBO enables a versatile multiplexing platform to perform qualitative and quantitative analysis of proteins and miRNAs in patient serum samples simultaneously. To the best of our knowledge, this is the first method to profile protein and miRNA biomarkers to diagnose DILI in a single-step assay.

By integrating Dynamic Chemical Labelling (DCL) with Luminex xMAP technology, it becomes feasible to analyse miRs through single- and multi-plex assays, as well as to simultaneously analyse proteins and miRs.

There is currently a need to investigate new biomarkers of acute liver injury (ALI) that are highly specific to the liver and capable of detecting early-stage liver damage. In this regard, circulating microRNAs (miRNAs), particularly microRNA122 (miRNA122), have recently been proposed as promising new candidate biomarkers. However, the quantification of miRNAs is not a standardized technique and presents several challenges that hinder its routine use. The aim of this work is to validate the innovative Dynamic Chemical Labeling (DCL) PCR-Free technology for its ability to detect miRNA122 in patient samples using Luminex xMAP platforms. The DCL PCR-Free technology was used to directly measure and monitor miRNA122 levels in serum samples from patients with ALI. Patients were monitored throughout the recovery process from liver injury, from the time of detection and for up to 30 days afterwards, with follow-up over three separate visits. The results demonstrate the robustness of the method, with sensitivity of 3.36% and precision of 99.80%, and show a high correlation between miRNA122 and traditional liver injury markers such as ALT (r = 0.8150, p = 0.0001), AST (r = 0.7895, p = 0.0002) and TBL (r = 0.2646, p = 0.3406) throughout the ALI recovery process. In conclusion, measuring miRNA122 levels using the DCL PCR-Free method provides a distinctive approach, not only for diagnosing ALI patients but also for effectively monitoring disease progression, tracking liver recovery, and evaluating treatment effectiveness.

MicroRNA‐122 (miR‐122) is a sensitive biomarker for paracetamol‐induced liver injury. This study assessed the diagnostic performance of dynamic chemical labelling (DCL) combined with an immunoassay platform to quantify miR‐122 in human serum, as a proof‐of‐concept for hospital use. This study used serum samples from 19 healthy individuals and 19 patients with paracetamol drug‐induced liver injury (DILI) collected in previous studies. miR‐122 was labelled with a biotinylated SMART‐Base and quantified using a multiplex, bead‐based, fluorescence detection system. The median miR‐122 concentration was significantly elevated at 233.4 pg/mL (IQR: 74.5–363.4) compared to 15.2 pg/mL (IQR: 9.4–24) in the control group. The Mann–Whitney U test showed significant group differences ( p < 0.0001). ROC analysis yielded an area under the curve (AUC) of 0.96, with a 29.7 pg/mL cut‐off providing 92.6% sensitivity (95% CI: 76.6%–98.7%) and 89.4% specificity (95% CI: 68.6%–98.1%). miR‐122 moderately correlated with ALT ( r = 0.56, p = 0.03). DCL with fluorescence detection is a promising clinical tool for miR‐122 quantification.