Glomerular deposition of hepatitis B virus (HBV) antigens are observed in chronic HBsAg carriers with different glomerulonephritides yet the etiologic role of HBV remains uncertain. We examined the paraffin section of kidney biopsies from 40 chronic HBsAg carriers with membranous nephropathy (MGN), mesangiocapillary glomerulonephritis (MCGN) or IgA nephropathy (IgAN) for HBV DNA and HBV RNA using in situ hybridization (ISH). Glomerular HBV antigens were present in all biopsies by immunofluorescence. HBsAg or HBcAg mRNA was also studied in RNA extracted from frozen renal tissue using a two-step polymerase chain reaction (PCR) following reverse transcription (RT). HBcAg DNA was not easily detected with ISH alone, but was readily found in 31 biopsies (78%) following PCR. HBV DNA was detected mainly in the cytoplasm of proximal tubular epithelia but not in glomerular cells. HBsAg and/or HBcAg mRNA were detected by RT-PCR in extracted RNA from 13 biopsies (33%). The PCR findings were further confirmed by (a) Southern blot hybridization using a cloned HBV probe and (b) absence of PCR product following treating RNA with RNase or omitting the RT. It is plausible that HBV DNA in renal tubules represents endocytosis of HBV DNA in the urinary filtrate and the HBV RNA extracted from kidney biopsies could derive from infiltrating cells bearing HBV RNA. Hence, ISH with specific HBV core gene RNA probe was performed subsequently. HBcAg RNA, localized in the nuclei and cytoplasm of glomerular and tubular cells, was detected in 56%, 20%, and 36% of renal biopsies in chronic HBsAg carriers with MGN, MCGN, and IgAN, respectively. Our findings indicate the presence of viral transcription in glomerular cells and renal tubular epithelia, supporting an etiological role of HBV in some chronic HBsAg carriers who develop coexisting glomerulonephritides.