Pathological and molecular-based virological studies of TTV and s-TTV infection

TTV 和 s-TTV 感染的病理学和分子病毒学研究

基本信息

批准号：
12670223
负责人：
ABE Kenji
金额：
$ 0.77万
依托单位：
National Institute of Infectious Diseases
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670223/
关键词：
TTV Simian TTV (s-TTV)TTV DNA TTV particle Molecular epidemiology ヒトDNAウイルス Simian TTV(s-TTV)肝疾患ウイルス粒子 Simian TTV s-TTV 霊長類チシパンジー PCR

项目摘要

1) Detection of TTV DNA in body fluids :TTV DNA was detected in the saliva of 7 (7/10 ; 70%) individuals and the semen of 6 (6/10 : 60%) individuals, respectively. In addition, TTV DNA was also detected frequently (8/16 ; 50%) in bile juice, suggesting a fecal-oral TTV transmission route.2) Observation of viral particles :Under the electron microscopic examination, viral-like particles (VLP) with 20-30nm in diameter were found in bile juice from a patient infected with TTV. VLPs were found in fractions with a density of 1.24-1.28/ml in continuous gradient of CsCl.3) Infection of simian TTV (s-TTV) to humans :s-TTV DNA was determined by nested PCR using s-TTV-specific primers designed from untranslated region of s-TTV genome. s-TTV DNA sequence was detected in 3 of 200 (1.5%) healthy adults. On the other hand, s-TTV DNA was detected in 18 of 260 (6.9%) Japanese patients with liver diseases (chronic hepatitis 10/217 ; 4.6%, cirrhosis 4/28 ; 142%, hepatocellular carcinoma 4/12; 33.3%). 55.5% (10/18) of patients were positive for s-TTV alone. s-TTV DNA was detected in 10/151 (6.6%) patients with liver diseases of unknown etiology. Nucleotide sequence analyses in 20 patients confirmed that these PCR products were derived from s-TTV genome sequences and nearly 85% identical to those of s-TTV prototypes from chimpanzees. Phylogenetic analysis demonstrated that all s-TTV isolates from humans were distinguished clearly from the human TTV isolates. Furthermore, s-TTV in humans was classified into 2 different genotypes as well as simians.

1)体液中TTV DNA的检测：分别在7名(7/10；70%)个体的唾液和6名(6/10：60%)个体的精液中检测到TTV DNA。此外，胆汁中也频繁检测到TTV DNA（8/16；50%），提示TTV存在粪口传播途径。2）病毒颗粒的观察：电镜下观察到病毒样颗粒（VLP）在感染TTV的患者的胆汁中发现了直径为20-30nm的颗粒。在 CsCl 连续梯度中，在密度为 1.24-1.28/ml 的级分中发现了 VLP。3) 猿 TTV (s-TTV) 对人类的感染：使用 s-TTV 特异性引物通过巢式 PCR 测定 s-TTV DNA从 s-TTV 基因组的非翻译区域设计。在 200 名健康成年人中，有 3 名 (1.5%) 检测到了 s-TTV DNA 序列。另一方面，在 260 名日本肝病患者中，有 18 名（6.9%）检测到 s-TTV DNA（慢性肝炎 10/217；4.6%，肝硬化 4/28；142%，肝细胞癌 4/12；33.3%））。 55.5% (10/18) 的患者仅 s-TTV 呈阳性。在 10/151 (6.6%) 患有不明原因肝病的患者中检测到 s-TTV DNA。对 20 名患者的核苷酸序列分析证实，这些 PCR 产物源自 s-TTV 基因组序列，与黑猩猩的 s-TTV 原型有近 85% 的一致性。系统发育分析表明，所有来自人类的 s-TTV 分离株均与人类 TTV 分离株有明显区别。此外，人类和猿猴中的 s-TTV 被分为 2 种不同的基因型。