首页> 外文期刊>The American Journal of Tropical Medicine and Hygiene >Development of a TaqMan(R) Minor Groove Binding Protein Assay for the Detection and Quantification of Crimean-Congo Hemorrhagic Fever Virus.
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Development of a TaqMan(R) Minor Groove Binding Protein Assay for the Detection and Quantification of Crimean-Congo Hemorrhagic Fever Virus.

机译:用于检测和定量克里米亚-刚果出血热病毒的TaqMan(R)小沟结合蛋白测定法的开发。

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摘要

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus of the genus Nairovirus and the family Bunyaviridae. It is a negative-strand RNA virus comprised of small (S), medium (M), and large (L) genome segments. The S segment encodes for nucleocapsid protein, the M segment codes for envelope glycoproteins (Gn and Gc), and the L segment codes for the RNA-dependent RNA polymerase. Currently, there are a limited number of methods for rapidly diagnosing CCHFV infections. We developed a real-time, reverse transcription-polymerase chain reaction assay for the rapid detection of CCHFV by using the TaqMan((R))-minor groove binding protein probe technology. The primers and probes were designed to amplify and detect a region in the S segment of CCHFV that is conserved across multiple strains. The limit of detection of the assay was 10 genome copies of RNA. This primer and probe set was specific to 18 strains of CCHFV tested and did not cross-react with either a DNA panel of 78 organisms or a panel of 28 diverse RNA viruses. This will rapidly and specifically detect CCHFV, and it has been used to detect CCHFV infection in samples from humans, animals, and ticks.
机译:克里米亚-刚果出血热病毒(CCHFV)是奈洛病毒属和Bunyaviridae家族的壁虱传播病毒。它是由小(S),中(M)和大(L)基因组片段组成的负链RNA病毒。 S段编码核衣壳蛋白,M段编码包膜糖蛋白(Gn和Gc),L段编码RNA依赖性RNA聚合酶。当前,用于快速诊断CCHFV感染的方法数量有限。我们开发了实时逆转录聚合酶链反应测定法,以通过使用TaqMan(-)-小沟结合蛋白探针技术快速检测CCHFV。设计引物和探针以扩增和检测CCHFV S段中跨多个菌株保守的区域。该测定的检测极限是RNA的10个基因组拷贝。该引物和探针组对测试的18种CCHFV菌株具有特异性,并且不会与78种生物体的DNA面板或28种不同RNA病毒的面板交叉反应。这将快速而特异性地检测CCHFV,并且已被用于检测人,动物和壁虱样品中的CCHFV感染。

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