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[粪便中一种非甲非乙型肝炎相关物质——一种部分双链环状DNA的鉴定与克隆]

[A hepatitis non-A, non-B-associated substance in the feces--identification and cloning of a partially double-stranded circular DNA].

作者信息

Burckhardt J, Seelig R, Calvo-Riera F, Seelig H P

机构信息

Institut für Immunologie und experimentelle Pathologie GmbH, Karlsruhe.

出版信息

Immun Infekt. 1988 Jun;16(3):91-6.

PMID:2841237
Abstract

By means of a radioimmunoassay a substance excreted in feces could be detected in patients with hepatitis non-A,non-B (HNANB). Feces extracts of patients with sporadic and posttransfusion HNANB as well as of healthy persons were precipitated with PEG, digested with RNase and DNase and separated on CsCl. In HNANB-patients a RIA-positive material with a density of 1.3 g/ml CsCl could be detected which contained a partially double-stranded circular DNA. Cloning of this DNA in lambda-phase resulted in DNA of about 5 Kb, which hybridized with feces DNA under stringent conditions. The 5 Kb-DNA were mapped with different restriction enzymes. A 1.5 Kb EcoRi-fragment cross-hybridizes with HBV-DNA. No hybridization and sequence homologies were found with human, viral and procaryotic DNA as well as with plasmid and phage DNA (data base EMBL, Heidelberg). It is assumed that the DNA excreted in feces of HNANB-patients represents a viral genome not detected so far.

摘要

通过放射免疫分析法,在非甲非乙型肝炎(HNANB)患者的粪便中可检测到一种物质。将散发性和输血后HNANB患者以及健康人的粪便提取物用聚乙二醇(PEG)沉淀,用核糖核酸酶(RNase)和脱氧核糖核酸酶(DNase)消化,然后在氯化铯(CsCl)中进行分离。在HNANB患者中,可检测到一种密度为1.3g/ml CsCl的放射免疫分析阳性物质,其含有部分双链环状DNA。将该DNA克隆到λ噬菌体中得到约5kb的DNA,在严格条件下该DNA与粪便DNA杂交。用不同的限制性内切酶对5kb的DNA进行图谱分析。一个1.5kb的EcoRⅠ片段与乙肝病毒(HBV)DNA交叉杂交。未发现与人类、病毒和原核生物DNA以及质粒和噬菌体DNA(数据库EMBL,海德堡)有杂交和序列同源性。推测HNANB患者粪便中排出的DNA代表一种迄今未检测到的病毒基因组。

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