Validation of combo ichroma as a reliable concentration-based alternative for AST and ALT measurement in liver disease monitoring

Background

Traditional assays for aspartate aminotransferase (AST) and alanine aminotransferase (ALT) measure enzymatic activity, which degrades during long-term frozen storage, threatening the accuracy of the assay. Instead, Combo ichroma (CI), a fluorescence immunoassay that quantifies AST and ALT concentrations, is a robust alternative for retrospective and point-of-care liver function testing, free from the influence of long-term storage.

Methods

Serum samples from 256 individuals (controls and patients with hepatitis, cirrhosis, or liver cancer) were collected and stored at −80 °C for an average of three years. AST and ALT were measured using CI, the 7180 clinical analyzer (HT), and Atellica CH 930 analyzer performed immediately after collection (HL). Correlations between AST, ALT, and AST/ALT ratios were analyzed. Random Forest Regression (RFR) models using CI or HT data were developed to predict HL-derived AST/ALT ratios.

Results

CI-AST showed strong correlation with HL-AST across all groups (R2 > 0.95), outperforming HT-AST, especially in controls (R2 = 0.58). CI-ALT moderately correlated with HL-ALT (R2 = 0.87), surpassing HT-ALT (R2 = 0.71). AST/ALT ratios varied across methods due to ALT variability, but RFR using CI data accurately predicted HL ratios (R2 = 0.85–0.91). Subgroup analysis confirmed CI’s superior concordance across etiologies.

Conclusions

CI enables activity-independent, reliable measurement of AST and ALT even after extended storage, outperforming enzymatic assays in precision and correlation. Its simplicity, and compatibility with machine learning models position CI as a promising tool for liver enzyme diagnostics in both clinical and resource-limited settings.