Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most prevalent liver disorder in Western countries, with approximately 20%-30% of the MASLD patients progressing to severe stages. There is an urgent need for noninvasive, cost-effective, widely accessible, and precise biomarkers to evaluate liver steatosis. This study aims to assess and compare the diagnostic performance of a novel reference frequency method-based ultrasound attenuation coefficient estimation (ACE) in both fundamental (RFM-ACE-FI) and harmonic (RFM-ACE-HI) imaging for detecting and grading liver steatosis. An Institutional Review Board-approved prospective study was carried out between December 2018 and October 2022. A total number of 130 subjects were enrolled in the study. The correlation between RFM-ACE-HI values and magnetic resonance imaging proton density fat fraction (MRI-PDFF), as well as between RFM-ACE-FI values and MRI-PDFF were calculated. The diagnostic performance of RFM-ACE-FI and RFM-ACE-HI was evaluated using receiver operating characteristic (ROC) curve analysis, as compared to MRI-PDFF. The reproducibility of RFM-ACE-HI was assessed by interobserver agreement between two sonographers. A strong correlation was observed between RFM-ACE-HI and MRI-PDFF, with R = 0.88 (95% confidence interval [CI]: 0.83-0.92; P < .001), while the correlation between RFM-ACE-FI and MRI-PDFF was R = 0.65 (95% CI: 0.50-0.76; P < .001). The area under the ROC (AUROC) curve for RFM-ACE-HI in staging liver steatosis grades of S ≥ 1 and S ≥ 2 was 0.97 (95% CI: 0.91-0.99; P < .001) and 0.98 (95% CI: 0.93-1.00; P < .001), respectively, and 0.76 (95% CI: 0.65-0.85) and 0.80 (95% CI: 0.70-0.88) for RFM-ACE-FI, respectively. Great reproducibility was achieved for RFM-ACE-HI, with an interobserver agreement of R = 0.97 (95% CI: 0.94-0.99; P < .001). The novel RFM-ACE-HI method offered high liver steatosis diagnostic accuracy and reproducibility, which has important clinical implications for early disease intervention and treatment evaluation.
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