论文部分内容阅读
本文报道ＨＢＶ基因扩增产物的限制性酶切分析和寡核苷酸探针杂交鉴定、应用血清ＨＢＶＤＮＡ的ＰＣＲ扩增产物制备基因探针和单项抗ＨＢｃ阳性血清的ＨＢＶ基因扩增。ＰＣＲ用ａｄｒ、ａｄｗ、ａｙｗ３种亚型ＨＢＶ的Ｃ基因区共有序列为内外两对引物分单次或套式扩增，产物为６６ａｂｐ或４２８ｂｐ，两者序列中段共含－ＢｇＩⅡ酶切点并与一寡核苷酸探针同源。４２８ｂｐ产物被用以缺口平移标记３２Ｐ探针作斑点和转印杂交检测。结合分子杂交的ＰＣＲ结果从单项抗ＨＢｃ阳性的８／４２例慢性肝炎和２／１２例无症状受测者血清中检出ＨＢＶＤＮＡ，证实病毒低度复制和传染性。
This article reports the restriction analysis of HBV gene amplification products and oligonucleotide probe hybridization identification, gene amplification of HBVDNA PCR amplification products and single anti-HBc-positive serum HBV gene amplification. PCR with adr, adw, ayw three subtypes HBV C gene region of the consensus sequence for both internal and external pairs of primers in a single or nested amplification, the product was 66abp or 428bp, the middle of the sequence containing two-BgI Ⅱ enzyme cutting point and An oligonucleotide probe is homologous. The 428 bp product was spot-blot hybridized with a gap-shift labeled 32P probe. Based on the PCR results of molecular hybridization, HBVDNA was detected in sera from 8/42 chronic hepatitis and 2/12 asymptomatic subjects with anti-HBc positive, confirming the low replication and infectivity of the virus.