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[基于噬菌体M13构建用于与甲型肝炎病毒RNA杂交的探针]

[Construction of probes for hybridization with hepatitis A virus RNA based on phage M13].

作者信息

Karginov V A, Zelenin S M, Bondar' A A, Nikolaeva E S, Naumov V A

出版信息

Mol Gen Mikrobiol Virusol. 1988 Jul(7):39-42.

PMID:2848193
Abstract

Three fragments of a DNA copy complementary to tre hepatitis A virus RNA have been isolated from the pHAV-VPI-22 plasmid and cloned in M 13 mp 8 vector. The 140, 250 and 390 b. p. fragments corresponded to the viral genome fragment coding for VP1 protein. Single-stranded DNAs of the hybrid phages with 250 b. p. and 390 b. p. inserts have been used for radioactive probe synthesis. The specificity of the probes for viral RNA was confirmed in hybridization with hepatitis A virus RNA purified from the virus-infected cell culture. The hybridization was shown to detect up to 10(-12)-10(-13) g of the virus. A principal possibility to use the probes in diagnostic purposes was demonstrated by the virus detection in blood samples obtained from patients with hepatitis A infection.

摘要

已从pHAV - VPI - 22质粒中分离出与甲型肝炎病毒RNA互补的DNA拷贝的三个片段,并克隆到M 13 mp 8载体中。140、250和390个碱基对的片段对应于编码VP1蛋白的病毒基因组片段。具有250个碱基对和390个碱基对插入片段的杂交噬菌体的单链DNA已用于放射性探针合成。在与从病毒感染细胞培养物中纯化的甲型肝炎病毒RNA杂交时,证实了探针对病毒RNA的特异性。杂交显示可检测到低至10(-12)-10(-13)克的病毒。通过在甲型肝炎感染患者的血液样本中检测病毒,证明了将这些探针用于诊断目的的主要可能性。

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