Hbeag

Laboratory tests of HBV serologic markers included HBsAg, anti-HBs, HBeAg, and anti-HBe (Abbott Laboratories, Abbott Park, IL, USA). The eGFR was calculated by the CKD Epidemiology Collaboration (CKD-EPI) equation. The measurement of urine protein included UPCR, UACR, and urine β2-microglobulin-to-creatinine ratio (AnGene Biotechnology Co., Ltd.). Anti-HDV IgG was determined by competitive enzyme immunoassay (Dia.Pro Diagnostic Bioprobes, Srl, Milan, Italy). For participants with a positive anti-HDV IgG, HDV RNA was determined with the use of in-house real-time PCR assay. In brief, the HDV nucleic acid was first extracted from 500-μL plasma using the QIAamp UltraSens virus kit (Qiagen, Hilden, Germany), and cDNA was reverse transcribed from 10 μL eluted RNA using random hexamer primer. The HDV RNA was then determined using a commercialized HDV quantification primer probe set (TIB LightMix kit) (38 (link), 39 (link)) with a detection limit of 10 copies/reaction. The amplification target of HDV was 113 bp on HDV delta antigen.
BMD assessment was performed in participants enrolled at four participating hospitals using dual-energy X-ray absorptiometry (Lunar Prodigy; GE Healthcare, Belgium). Osteoporosis and osteopenia were defined by BMD T-score according to WHO criteria (40 (link)).

The lipid profile (TC, TGs, and HDL-C) and alanine aminotransaminase (ALT), fasting glucose (Glu), HBsAg, HBeAg (Abbott Laboratories, North Chicago, IL), HBV-DNA (Digene Corp., Gaithersburg, MD), HCV Ab (Abbott Laboratories, Chicago, IL), and HCV-RNA levels (Roche Diagnostics, Tokyo, Japan) and HCV genotypes (Roche Diagnostics) were assessed at the clinical pathology or liver research laboratory of the authors’ affiliated hospital using routine automated techniques.