Detection of HBV replicated intermediates in the liver of HBV(ayw) transgenic mice

Objective To establish highly sensitive and stable chemiluminescent Southern blotting system,and to detect the replication levels of HBV DNA in liver,kidney,spleen and muscles of HBV(ayw) transgenic mice.Methods The HBV probe was labeled with digoxigenin with PCR,and the sensitivity and reproducibility of hybridization system were detected by pTHBV1047.The total genomic DNA was extracted from the tissues of HBV transgenic mice and digested with Hind Ⅲ for 18 hours.After eletrophoresis,the digested DNA was transferred to nylon membrane.The replicated intermediates in the liver and other tissues of HBV transgenic mice were detected by chemiluminescent hybridization Southern blotting system.Results The sensitivity of the detection system labeled with digoxigenin was about 0.1pg,and the hybridization system was stable and reliable.Both the integrated transgene of HBV DNA and the HBV replicated intermediates were detected in the liver of HBV transgenic mice,but the level of HBV replication was lower than that in HepG2.2.15 cell line.HBV DNA replication also existed in kidney,spleen and muscles,but the replicated levels in these tissues were lower than that in the liver.Conclusions The 27th generation of HBV(ayw) transgenic mice is replicated in HBV transgenic mice with HBV gene stably integrated in the genome thereof.