Improvement of HBV DNAextraction method from human breast milk

Abstract

：Objective To investigatethe pre-tretment influence of human breast milk specimens on the HBV DNA quantitativedetection and optimize the methods of HBV DNA extraction from human breast milkspecimens.Methods Different methods including DNA kit,Alkaline lysis,acidolysis,anhydrousethanol and deepfreeze were used to extract HBV DNA from human breast milk specimens,withdifferent HBV DNA load,and HBV DNA was quantitatively detected by real-time PCR.ResultsDNA kit,Alkaline lysis,deep freeze were equally efficient to extract HBV DNA,with the rateof positive at 100%,when the human breast milk contained HBV DNA of>10~4 IU/mL.Moreovertheir coin-cidence rate of quantitative result was 100% in HBV DNA positive serum dilutedby human breast milk and HBV DNA negative serum.While,other two methods led a negativeresult.However,only alkaline lysis(NaOH concentration was0.4%-1.0%)achieved a positiveyield when the HBV DNA load of human breast milk specimens <10~4 IU/mL,and maintainedthe same coincidence rote of quantitative result at 100%.In contract,neither the otherfour pre-treatment methods gave a positive result.Conclunsions Alkaline lysis method canpotentially adapted to clinical medicine for HBV DNA extraction from human breast milkspecimens,with the NaOH concentration at 0.4%-1.0%.