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聚合酶链反应技术结合酶切杂交研究单项抗乙型肝炎病毒核心抗原阳性者的乙型肝炎病毒感染 被引量:4

Amplification of Hepatitis B Virus(HBV) DNA by Polymerase Chain Reaction(PCR) in Serum which is Solely Positive to Anti-hepatitis B Core Antigen(HBc)
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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
关键词 乙型肝炎病毒 核心抗原 DNA 聚合酶链反应 抗体 Polymerase chain reaction Hepatitis B virus Deoxyribonucleic acid Anti-HBc
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