DETECTION OF HEPATITIS-B VIRUS-DNA IN SERUM SAMPLES VIA NESTED PCR AND MALDI-TOF MASS-SPECTROMETRY

In a blind study, nested polymerase chain reaction (PCR) was performed with control DNA and DNA preparations from serum samples of six patie nts. The detection limit was determined to be 100 molecules of templat e in 1 mi of serum. Hepatitis B virus (HBV) related products of nested PCR were purified by ultrafiltration and immobilisation on streptavid in coated magnetic beads. The immobilized PCR products were denatured from the beads and analyzed via matrix assisted laser desorption/ionis ation time-of-flight (MALDI-TOF) mass spectrometry. The results of MAL DI-TOF MS analysis were in agreement with the results obtained by poly acrylamide gel electrophoresis (PAGE) and with the data obtained by se rological analysis. The detection strategy introduced here has a high potential for automation and represents a fast and reliable method of detection for HBV DNA in serum without the need for time consuming gel electrophoresis and labeling or hybridization procedures.