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利用泛病毒重测序微阵列技术重新研究在欧洲流行的新兴汉坦病毒的遗传多样性。

Revisiting the genetic diversity of emerging hantaviruses circulating in Europe using a pan-viral resequencing microarray.

机构信息

Institut Pasteur, Antiviral Strategies Unit, Department of Virology, Paris, France.

Institut Pasteur, Unit of Epidemiology and Physiopathology of Oncogenic Viruses, CNRS, UMR 3569, Department of Virology, Paris, France.

出版信息

Sci Rep. 2019 Aug 27;9(1):12404. doi: 10.1038/s41598-019-47508-7.

Abstract

Hantaviruses are zoonotic agents transmitted from small mammals, mainly rodents, to humans, where they provoke diseases such as Hemorrhagic fever with Renal Syndrome (HFRS) and its mild form, Nephropathia Epidemica (NE), or Hantavirus Cardio-Pulmonary Syndrome (HCPS). Hantaviruses are spread worldwide and monitoring animal reservoirs is of primary importance to control the zoonotic risk. Here, we describe the development of a pan-viral resequencing microarray (PathogenID v3.0) able to explore the genetic diversity of rodent-borne hantaviruses endemic in Europe. Among about 800 sequences tiled on the microarray, 52 correspond to a tight molecular sieve of hantavirus probes covering a large genetic landscape. RNAs from infected animal tissues or from laboratory strains have been reverse transcribed, amplified, then hybridized to the microarray. A classical BLASTN analysis applied to the sequence delivered through the microarray allows to identify the hantavirus species up to the exact geographical variant present in the tested samples. Geographical variants of the most common European hantaviruses from France, Germany, Slovenia and Finland, such as Puumala virus, Dobrava virus and Tula virus, were genetically discriminated. Furthermore, we precisely characterized geographical variants still unknown when the chip was conceived, such as Seoul virus isolates, recently emerged in France and the United Kingdom.

摘要

汉坦病毒是一种从小型哺乳动物(主要是啮齿类动物)传播给人类的人畜共患病原体,可引起肾综合征出血热(HFRS)及其轻度形式流行性肾病(NE),或汉坦病毒心肺综合征(HCPS)等疾病。汉坦病毒在全球范围内传播,监测动物宿主对于控制人畜共患病风险至关重要。在这里,我们描述了一种通用病毒重测序微阵列(PathogenID v3.0)的开发,该微阵列能够探索在欧洲流行的啮齿动物携带汉坦病毒的遗传多样性。在微阵列上约 800 个序列中,有 52 个对应于汉坦病毒探针的紧密分子筛,涵盖了广泛的遗传景观。感染动物组织或实验室菌株的 RNA 已被反转录、扩增,然后与微阵列杂交。通过微阵列提供的序列进行经典的 BLASTN 分析可识别汉坦病毒的物种,直至测试样本中存在的精确地理变异。对来自法国、德国、斯洛文尼亚和芬兰的最常见的欧洲汉坦病毒的地理变异体,如普马拉病毒、多布拉瓦病毒和图拉病毒进行了遗传区分。此外,我们还精确地描述了芯片设计时仍未知的地理变异体,如首尔病毒分离株,最近在法国和英国出现。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/13d5/6712034/82dfada2ec51/41598_2019_47508_Fig1_HTML.jpg

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