Thiers V, Nakajima E, Kremsdorf D, Mack D, Schellekens H, Driss F, Goudeau A, Wands J, Sninsky J, Tiollais P
Unit of Recombination and Genetic Expression, Institute Pasteur, Paris, France.
Lancet. 1988 Dec 3;2(8623):1273-6. doi: 10.1016/s0140-6736(88)92891-7.
The polymerase chain reaction (PCR) was used to identify and characterise serum HBV DNA sequences in 3 patients negative for all HBV serological markers. HBsAg determinants were detected in 1 individual by monoclonal anti-HBsAg immunoradiometric assay. By use of sets of primers on the S and pre-S parts of the HBV genome the presence of HBV DNA was demonstrated in the serum of all 3 patients. Inoculation of human sera to 2 chimpanzees induced acute hepatitis in both animals; 1 became positive for HBsAg and anti-HBcAg and the other only for anti-HBsAg. Cloning of DNA sequences from viral isolates from 1 chimpanzee and 1 patient was accomplished after amplification of the 3' region of the S gene. Comparison of the partial nucleotide sequence with that of known HBV subtypes showed 0 and 1 point mutation, respectively, in the highly conserved 3' end of the S gene. Therefore the results show that PCR with HBV primers may unambigously identify HBV infectious particles among non-A, non-B viruses and is a potentially useful diagnostic test for detection of HBV DNA sequences in serum.
采用聚合酶链反应(PCR)对3例所有乙肝病毒血清学标志物均为阴性的患者血清中的乙肝病毒(HBV)DNA序列进行鉴定和特征分析。通过单克隆抗-HBsAg免疫放射分析在1例个体中检测到HBsAg决定簇。利用针对HBV基因组S区和前S区的引物组,证实所有3例患者血清中均存在HBV DNA。将人血清接种到2只黑猩猩体内,两只动物均诱发了急性肝炎;1只黑猩猩HBsAg和抗-HBcAg呈阳性,另一只仅抗-HBsAg呈阳性。在扩增S基因3'区域后,完成了对1只黑猩猩和1例患者病毒分离株DNA序列的克隆。将部分核苷酸序列与已知HBV亚型的序列进行比较,结果显示在S基因高度保守的3'端分别有0个和1个点突变。因此,结果表明,使用HBV引物的PCR可明确鉴定非甲非乙型病毒中的HBV感染颗粒,并且是检测血清中HBV DNA序列的一种潜在有用的诊断试验。