Novel DNA sequence isolated from blood donors with high transaminase levels
Novel DNA sequence isolated from blood donors with high transaminase levels
复制标题
从转氨酶水平较高的献血者中分离出的新 DNA 序列
DOI：
10.1111/j.1872-034x.2011.00848.x
复制
发表时间：
2011
期刊：
影响因子：
--
通讯作者：
et al
中科院分区：
文献类型：
--
作者：
Sato;et al
目的：在日本，10%-20%的急性肝炎病例的病因尚不清楚。本研究旨在验证引起非甲-戊型肝炎的病原体。方法：利用RNA病毒解旋酶保守区构建的引物，对500例丙氨酸氨基转移酶(ALT)升高的献血员血清标本进行聚合酶链式反应筛查。结果：4份血样中均含有一个新的 序列，命名为Kis-V，对限制性内切酶SalI和BstXI敏感。滚环扩增产生过量的Kis-V DNA。在蔗糖密度梯度超速离心法中，Kis-V的密度为1.158-g/cm~3。洗涤剂处理使Kis-V密度增加，人白细胞DNA未见Kis-V DNA扩增。序列筛选表明Kis-V包含在一个大小为30-50 nm的颗粒中，电子分析显示Kis-V含有13个潜在基因，没有一个与已报道的病毒蛋白同源。其中一个基因与DNA聚合酶结构域相似。发现了强烈的转录启动信号和CpG岛。Kis-V的核苷酸组成具有环状DNA基因组的特征，包括复制起点和末端。在一项初步研究中，在ALT水平升高的戊型肝炎病毒抗体阳性个体中经常发现Kis-V。结论：从ALT水平升高的献血员中分离到新的Kis-V序列。有人认为Kis-V是一种双链环状DNA基因组，来源于一类新的囊膜病毒。
Aim:In Japan, the etiology of 10–20% of cases of acute hepatitis remains unclarified. This study was conducted to verify the agent causing non‐A–E hepatitis.Methods:Serum samples from 500 blood donors with elevated alanine aminotransferase (ALT) levels were screened by polymerase chain reaction using primers constructed from conserved areas of RNA virus helicase. The sequence obtained was investigated for viral properties.Results:Four blood samples were found to contain a novel DNA sequence of 9496 bp, which was designated KIs‐V. KIs‐V was sensitive to the restriction enzymeSalI andBstXI. Rolling‐circle amplification produced an excessive amount of KIs‐V DNA. In sucrose density gradient ultracentrifugation, KIs‐V banded at a 1.158‐g/cm3density. Detergent treatment increased the density of KIs‐V. There was no KIs‐V DNA amplification from human leukocyte DNA. Serial filtration suggested that KIs‐V was included in a 30–50‐nm size particle.In silicoanalysis revealed that KIs‐V contained 13 potential genes, none of which showed homology to any viral proteins reported. One gene showed similarity to a DNA polymerase domain. Strong signals for transcription initiation and a CpG island were identified. The nucleotide composition of KIs‐V showed a characteristic feature of circular DNA genomes that contain a replication origin and a terminus. In a preliminary study, KIs‐V was frequently identified among hepatitis E virus antibody positive individuals with elevated ALT levels.Conclusion:A new sequence KIs‐V was isolated from blood donors with elevated ALT levels. It was suggested that KIs‐V is a double‐stranded circular DNA genome derived from a novel category of enveloped viruses.