Since publication of the above referenced manuscript, it has come to the authors' attention that the PCO (sense) primer described in Table I lies upstream of the start site of the viral pregenomic RNA. Accordingly, this primer cannot amplify reversed transcribed viral RNA as illustrated in Fig. 7. The signal illustrated in that figure must have been generated, therefore, by a hemi-nested PCR reaction involving the COR, PCN, and NCO primers shown in Table II. The authors have recently confirmed the presence of HBV RNA sequences in the PBMC of case 4, as shown in Fig. 7, using PCN (sense)-COR (antisense) primers for initial amplification followed by 1898 (sense)-NCO (antisense) nested amplification. The coordinates and sequence of all primers are shown in Table II except 1898 sense, which represents position 1898-1917 in HBV and contains the sequence 5'-CATGGACATCGACCCTTATA-3'. PBMC from case 2 are not available for retesting; however, the authors have reproduced these results in PBMC RNA derived from two patients with chronic HBV infection using the same series of primers, as well as with primers covering the envelope and X regions of the viral genome.

Contrary to current opinion, the disappearance of hepatitis B surface antigen (HBsAg) from the serum, the development of anti-HBs antibodies, and normalization of liver function may not reflect complete virological recovery from acute hepatitis B virus (HBV) infection. By using the polymerase chain reaction (PCR), in the current study we demonstrate long-term persistence of HBV DNA in the serum and peripheral blood mononuclear cells (PBMC) of four patients for up to 70 mo after complete clinical, biochemical, and serological recovery from acute viral hepatitis. Serum HBV DNA reactivity co-sedimented with HBsAg in sucrose gradients, and it displayed the size and density characteristics of naked core particles and intact HBV virions, presumably contained within circulating immune complexes in these anti-HBs antibody-positive sera. HBV DNA was also present in PBMC in late convalescent samples from all four patients, and HBV RNA was detected in late convalescent phase PBMC in two of these patients. These results suggest that HBV DNA, and possibly HBV virions, can be present in the serum, and that the viral genome can persist in a transcriptionally active form in PBMC for > 5 yr after complete clinical and serological recovery from acute viral hepatitis. (J.