Gomes S A, Yoshida C F, Niel C
Department of Virology, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil.
Acta Virol. 1996 Jun;40(3):133-8.
The presence of hepatitis B virus (HBV) DNA was investigated by polymerase chain reaction (PCR) in the serum of twenty Brazilian blood donors. All sera were negative for hepatitis B surface antigen (HBsAg), 17 of them presented antibodies to the hepatitis B core antigen (anti-HBc) as the unique serological marker of HBV infection, and 3 were positive for antibodies to HBsAg (anti-HBs) and anti-HBc. PCR assays were carried out using different pairs of oligonucleotides designed from conserved sequences of C, pre-S and S regions of the HBV genome. First, all oligonucleotide pairs were tested in PCR using plasmids carrying HBV genome from ayw or adw strains as templates. One-round PCR assays were able to detect 100-25,000 molecules of plasmid DNA, depending on the oligonucleotide pair, while semi-nested PCR assays detected 10-1000 molecules. The frequency of HBV DNA-positive results with HBsAg-sera varied from 0% to 50% depending upon the PCR assay. The results indicated that a number of both isolated anti-HBc and anti-HBs+, anti-HBc+ samples contained HBV DNA at a very low concentration, neighboring the limit of detection.
采用聚合酶链反应（PCR）对20名巴西献血者血清中的乙型肝炎病毒（HBV）DNA进行了检测。所有血清的乙型肝炎表面抗原（HBsAg）均为阴性，其中17人仅以乙型肝炎核心抗原抗体（抗-HBc）作为HBV感染的唯一血清学标志物，3人乙型肝炎表面抗原抗体（抗-HBs）和抗-HBc呈阳性。使用根据HBV基因组C区、前S区和S区保守序列设计的不同寡核苷酸对进行PCR检测。首先，以携带ayw或adw株HBV基因组的质粒为模板，在PCR中对所有寡核苷酸对进行检测。一轮PCR检测能够检测到100 - 25,000个质粒DNA分子，具体取决于寡核苷酸对，而半巢式PCR检测能检测到10 - 1000个分子。根据PCR检测方法的不同，HBsAg阴性血清中HBV DNA阳性结果的频率在0%至50%之间。结果表明，一些单独的抗-HBc样本以及抗-HBs +、抗-HBc +样本中均含有极低浓度的HBV DNA，接近检测限。
