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利用RNA-DNA杂交技术鉴别蜱传脑炎病毒毒株

Differentiation of strains of tick-borne encephalitis virus by means of RNA-DNA hybridization.

作者信息

Shamanin V A, Pletnev A G, Rubin S G, Zlobin V I

机构信息

Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the Academy of Sciences of the U.S.S.R.

出版信息

J Gen Virol. 1990 Jul;71 ( Pt 7):1505-15. doi: 10.1099/0022-1317-71-7-1505.

Abstract

Cloned cDNA and synthetic deoxyoligonucleotides, complementary to various parts of the genomic RNA of tick-borne encephalitis virus (TBEV), strain Sofjin, were used to distinguish between strains of TBEV and other flaviviruses. The cDNA probe hybridized with strains of TBEV and related flaviviruses of the TBE complex except for Powassan virus, and it did not react with flaviviruses of the Japanese encephalitis and dengue subgroups. Viruses of the TBE complex and some strains of TBEV were differentiated from TBEV strain Sofjin by the thermal stability of RNA-DNA hybrids. Negishi and louping-ill viruses were the most closely related to TBEV strain Sofjin, among viruses of the TBE complex. Eight strains of TBEV isolated in different geographical areas from different sources were tested by dot-hybridization with 11 deoxyoligonucleotide probes. The probes revealed genetic variations among strains of TBEV. The pattern of hybridization correlated with the source of virus strains: TBEV strains isolated from TBE patients reacted with more probes than strains isolated from ticks. Within a group of epidemic strains of TBEV there was a correlation between the geographical area of isolation and similarity to TBEV strain Sofjin.

摘要

克隆的互补DNA(cDNA)和合成的脱氧寡核苷酸与蜱传脑炎病毒(TBEV)索夫金株基因组RNA的不同部分互补,用于区分TBEV的不同毒株以及其他黄病毒。该cDNA探针能与TBEV毒株以及TBE病毒复合群中除波瓦桑病毒外的相关黄病毒杂交,但不与日本脑炎和登革热亚群的黄病毒发生反应。通过RNA-DNA杂交体的热稳定性,可将TBE病毒复合群的病毒及部分TBEV毒株与TBEV索夫金株区分开来。在TBE病毒复合群的病毒中,根岸病毒和跳跃病病毒与TBEV索夫金株关系最为密切。用11种脱氧寡核苷酸探针通过点杂交对来自不同地理区域、不同来源的8株TBEV进行了检测。这些探针揭示了TBEV毒株之间的遗传差异。杂交模式与病毒毒株的来源相关:从TBE患者分离出的TBEV毒株比从蜱虫分离出的毒株能与更多探针发生反应。在一组TBEV流行毒株中,分离地区与和TBEV索夫金株的相似性之间存在相关性。

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