COMPARISON OF METHODS FOR EXTRACTION OF NUCLEIC-ACID FROM HEMOLYTIC SERUM FOR PCR AMPLIFICATION OF HEPATITIS-B VIRUS-DNA SEQUENCES

The sensitivity of PCR for the amplification of target nucleic acid se quences in clinical diagnostics may often be reduced due to the presen ce of inhibitory factors. Hemolytic serum contains a number of PCR inh ibitors, one of which is hemin. In this study we have found that conve ntional methods of DNA extraction were not sufficient for the removal of PCR-inhibitory compounds in hemolytic serum. We have therefore comp ared the efficiency of several commercial and noncommercial methods of nucleic acid purification from hemolytic serum samples prior to PCR a mplification. Separation with the QIAamp HCV kit, dialysis with Millip ore filters, and bovine serum albumin absorption were all found to be suitable extraction methods for eliminating inhibitors from hemolytic serum for PCR amplification. Using these methods we were able to detec t very low levels of hepatitis B virus DNA in hemolytic serum.