Garrison A R, Alakbarova S, Kulesh D A, Shezmukhamedova D, Khodjaev S, Endy T P, Paragas J
Virology Division and Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland, USA.
Am J Trop Med Hyg. 2007 Sep;77(3):514-20.
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.
克里米亚-刚果出血热病毒（CCHFV）是内罗毕病毒属、布尼亚病毒科的一种蜱传病毒。它是一种负链RNA病毒，由小（S）、中（M）和大（L）基因组片段组成。S片段编码核衣壳蛋白，M片段编码包膜糖蛋白（Gn和Gc），L片段编码RNA依赖性RNA聚合酶。目前，快速诊断CCHFV感染的方法有限。我们利用TaqMan®小沟结合蛋白探针技术开发了一种实时逆转录聚合酶链反应检测方法，用于快速检测CCHFV。引物和探针设计用于扩增和检测CCHFV S片段中多个菌株保守的区域。该检测方法的检测限为10个RNA基因组拷贝。该引物和探针组对测试的18株CCHFV具有特异性，与78种生物的DNA面板或28种不同RNA病毒的面板均无交叉反应。这将快速、特异性地检测CCHFV，并且已用于检测来自人类、动物和蜱的样本中的CCHFV感染。
