摘要
以聚合酶链反应(PCR)扩增单项抗乙型肝炎病毒核心抗原(HBc)阳性血清的乙型肝炎病毒(HBV)DNA。内外两对引物均选自adr、adw和ayw3种亚型HBV基因组C基因区的共有序列,扩增产物分别长428bP和664bP,共含一个BglⅡ酶切点,酶切后前者产生299bP和125bP、后者产生406bp和254bp各一个限制性片段。以另一条基因位置处于内引物对之间的寡核苷酸制备32P-5'末端标记的探针对之进行Southern转移杂交,鉴定了扩增产物的特异性。结果从单项抗HBc阳性的8/42例慢性肝炎和2/12倒无症状受测者血清中检出HBVDNA,提示现症低水平病毒复制和潜在的传染性。
Five oligonucleohdes nested in the C open reading frame(ORF) of the HBV genome and co-conserved by the subtpe adr, adw and ayw were synthesized 4 of which were pairedly used as primers in the polymerase chain reaction to amplify HBV DNA in serum which is solely positive to anti-HBc. The product of the outer primer pair is 664 bp in length, and of the inner 428 bp, both sharing a Bgl II restriction site in their sequences. After digestion with the enzyme, the 664 bp product yielded a 406 bp and a 254 bp fragment. and the 428 bp product yielded a 299 bp and a 125 bp. All of them were determined by Southern-blot hybridization that useing the other oligonucleotide as a probe, of which the sequence was nested between the inner pair and the 5' terminus labelled with 32P-dATP. Using these techniques. HBV DNA was detected in 8 of 42chronic hepatitis and 2 of 12 healthy carriers, indicating low-level viral replication as potential infectivity.
出处
《中华实验和临床病毒学杂志》
CSCD
1995年第4期318-321,共4页
Chinese Journal of Experimental and Clinical Virology