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从受感染人类血清和粪便中回收的TT病毒颗粒的可视化。

Visualization of TT virus particles recovered from the sera and feces of infected humans.

作者信息

Itoh Y, Takahashi M, Fukuda M, Shibayama T, Ishikawa T, Tsuda F, Tanaka T, Nishizawa T, Okamoto H

机构信息

Institute of Immunology, Tokyo, 112-0004, Japan.

出版信息

Biochem Biophys Res Commun. 2000 Dec 20;279(2):718-24. doi: 10.1006/bbrc.2000.4013.

Abstract

TT virus (TTV) has not yet been cultured or visualized. We attempted to recover and visualize TTV-associated particles from the serum samples and feces of infected humans. Serum samples were obtained from 7 human immunodeficiency virus (HIV)-infected patients. Three patients had a high TTV DNA titer (10(8) copies/ml), three had a low TTV DNA titer (10(2) copies/ml), and one was negative for TTV DNA. Fecal supernatant was obtained from a different TTV-infected subject. The serum samples were fractionated by high-performance liquid chromatography, and TTV DNA-rich fractions were subjected to floatation ultracentrifugation in cesium chloride. Virus-like particles, 30-32 nm in diameter, were found in the 1.31-1.33 g/cm(3) fractions from each of the three serum samples with high TTV DNA titer, but not in any fraction from the four serum samples that either were negative for TTV DNA or had low TTV DNA titer. The TTV particles formed aggregates of various sizes, and immunogold electron microscopy showed that they were bound to human immunoglobulin G. Similar virus-like particles with a diameter of 30-32 nm banding at 1.34-1.35 g/cm(3) were visualized in fecal supernatant with TTV genotype 1a by immune electron microscopy using human plasma containing TTV genotype 1a-specific antibody.

摘要

TTV病毒(TTV)尚未被培养或可视化。我们试图从受感染人类的血清样本和粪便中回收并可视化与TTV相关的颗粒。从7名感染人类免疫缺陷病毒(HIV)的患者中获取血清样本。3名患者的TTV DNA滴度较高(10⁸拷贝/毫升),3名患者的TTV DNA滴度较低(10²拷贝/毫升),1名患者的TTV DNA呈阴性。从另一名感染TTV的受试者中获取粪便上清液。血清样本通过高效液相色谱进行分级分离,富含TTV DNA的级分在氯化铯中进行漂浮超速离心。在三个TTV DNA滴度较高的血清样本的1.31 - 1.33克/立方厘米级分中发现了直径为30 - 32纳米的病毒样颗粒,但在四个TTV DNA呈阴性或TTV DNA滴度较低的血清样本的任何级分中均未发现。TTV颗粒形成了各种大小的聚集体,免疫金电子显微镜显示它们与人免疫球蛋白G结合。通过使用含有TTV基因型1a特异性抗体的人血浆进行免疫电子显微镜观察,在粪便上清液中也可视化到了直径为30 - 32纳米、在1.34 - 1.35克/立方厘米处出现条带的类似病毒样颗粒,其TTV基因型为1a。

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