LIGASE CHAIN-REACTION (LCR(R)) ASSAY FOR SEMIQUANTITATIVE DETECTION OF HBV DNA IN MONONUCLEAR LEUKOCYTES OF PATIENTS WITH CHRONIC HEPATITIS-B

A ligase chain reaction (LCR(R))-based approach to detect hepatitis B virus (HBV) DNA in peripheral blood mononuclear cells (PBMC) is descri bed. Using this new amplification technique, we determined semi-quanti tatively the amount of a short HBV S-gene fragment in PBMC lysates of 25 patients with different forms of chronic hepatitis (group A (n = 8) , hepatitis B s antigen (HBsAg)+/hepatitis B e antigen (HBeAg)+; group B (n = 9), HBsAg+/HBeAg-; group C (n = 8), HBsAg-/HBeAg-). The LCR re sults were compared with the findings obtained with polymerase chain r eaction (PCR) amplification of three distinct HBV gene regions (preS1/ 2, S and C) and related to the serological profiles of the patients. D epending on the primer pair used for PCR amplification, sensitivity of HBV LCR in PBMC was equivalent or slightly superior to PCR. The highe st positivity rate for HBV DNA was observed in the HBeAg+ and HBV DNA seropositive group (8/8) and was lower in the other patient groups B ( 4/9) and C (1/8). Interestingly, HBV gene sequences could also be dete cted in the lymphocytes of an HBsAg negative patient and in two patien ts from group B who were both negative for serum viral particles by PC R. The rapid LCR(R) procedure represents a reliable alternative to PCR for the sensitive detection of HBV DNA in PBMC samples. In combinatio n with the automated IMx(TM)-sgstem the new amplification technique ma y be routinely used for screening for HBV in whole blood samples and t hus may help to better evaluate the risk of HBV reinfection in liver t ransplant recipients.