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首页> 外文期刊>Journal of Clinical Microbiology >Highly Sensitive and Broadly Reactive Quantitative Reverse Transcription-PCR Assay for High-Throughput Detection of Rift Valley Fever Virus
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Highly Sensitive and Broadly Reactive Quantitative Reverse Transcription-PCR Assay for High-Throughput Detection of Rift Valley Fever Virus

机译：高灵敏度和广泛反应性的定量逆转录-PCR检测裂谷热病毒的高通量。

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Rift Valley fever (RVF) virus is a mosquito-borne virus associated with large-scale epizootics/epidemics throughout Africa and the Arabian peninsula. Virus infection can result in economically disastrous “abortion storms” and high newborn mortality in livestock. Human infections result in a flu-like illness, with 1 to 2% of patients developing severe complications, including encephalitis or hemorrhagic fever with high fatality rates. There is a critical need for a highly sensitive and specific molecular diagnostic assay capable of detecting the natural genetic spectrum of RVF viruses. We report here the establishment of a pan-RVF virus quantitative real-time reverse transcription-PCR assay with high analytical sensitivity (～5 RNA copies of in vitro-transcribed RNA/reaction or ～0.1 PFU of infectious virus/reaction) and efficiency (standard curve slope = ?3.66). Based on the alignments of the complete genome sequences of 40 ecologically and biologically diverse virus isolates collected over 56 years (1944 to 2000), the primer and probe annealing sites targeted in this assay are known to be located in highly conserved genomic regions. The performance of this assay relative to serologic assays is illustrated by testing of known RVF case materials obtained during the Saudi Arabia outbreak in 2000. Furthermore, analysis of acute-phase blood samples collected from human patients (25 nonfatal, 8 fatal) during that outbreak revealed that patient viremia at time of presentation at hospital may be a useful prognostic tool in determining patient outcome.
机译：裂谷热（RVF）病毒是一种蚊子传播的病毒，与整个非洲和阿拉伯半岛的大规模流行病/流行病有关。病毒感染会导致经济上灾难性的“人工流产风暴”和牲畜的高新生儿死亡率。人类感染会导致类似流感的疾病，其中1至2％的患者会出现严重的并发症，包括脑炎或出血热，死亡率高。迫切需要一种能够检测RVF病毒自然遗传谱的高度灵敏，特异的分子诊断方法。我们在此报告了建立具有高分析灵敏度（约5个RNA拷贝的体外转录RNA /反应或约0.1个PFU感染性病毒/反应）和高效性的pan-RVF病毒定量实时逆转录PCR检测方法的建立（标准曲线斜率= 3.66）。根据在56年间（1944年至2000年）收集的40种生态和生物多样性病毒分离株的完整基因组序列的比对，已知此测定法中靶向的引物和探针退火位点位于高度保守的基因组区域。通过检测2000年沙特阿拉伯暴发期间获得的已知RVF病例材料，可以说明该试验相对于血清学试验的性能。此外，分析了在该暴发期间从人类患者中收集的急性期血液样本（25例非致命，8例致命）揭示在医院就诊时患者病毒血症可能是确定患者预后的有用预后工具。

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