SYNTHESIS OF A NONRADIOACTIVE HEPATITIS-B VIRUS-DNA PROBE FROM HUMAN SERUM BY THE POLYMERASE CHAIN-REACTION

A method for synthesizing probes for detecting hepatitis B virus DNA i n serum was developed. It uses DNA extracted from the serum of an hepa titis B virus carrier as target, and digoxigenin-11-dUTP incorporated in DNA sequences during the polymerase chain reaction as tracer. Using a spot hybridization assay, the sensitivity and specificity of the di goxigenin-labelled DNA probe were compared with two standard hepatitis B virus DNA probes, synthesized with cloned hepatitis B virus DNA and labelled either with digoxigenin or P-32 by random priming. Data obta ined from the three methods showed an excellent correlation. Thus, hep atitis B virus DNA extracted from human serum and labelled by polymera se chain reaction may be considered a suitable alternative to cloned D NA. It provides reliable probes and eliminates the need for facilities and personnel dedicated to the manipulation of clones. These advantag es will allow a wider application of hepatitis B virus DNA testing in clinical practice.