Molecular detection of weaponized arenaviruses

描述（由申请人提供）：拟议研究的目的是设计、开发和评估分子检测方法，以便通过实时RT-PCR快速简便地检测生物恐怖主义背景下使用的武器化沙粒病毒（AV）。可灭活的AV包括CDC定义的A类病原体列表中分类的5种病毒（BSL-4因子），以及由突变和/或重组和/或重组株组成的所有潜在工程化AV。很少有基因组数据可用于AV，然而，在天然或实验产生的AV中已经观察到基因重组和重配。因此，了解所有AV的完整基因组序列是检测武器化AV的基石。为了区分BSL-4 AV与其他AV，其中至少四种在北美自然传播并可能感染人类，并且为了检测嵌合工程AV，确定BSL-4 AV以及BSL-2/3 AV的完整序列至关重要。该项目的实现需要在RNA病毒基因组学方面的丰富经验，人类病原体诊断检测设计/评估过程的专业知识，沙粒病毒遗传学的完美知识，BSL-4病原体的无可争议的工作经验，以及BSL-4实验室的即时可用性。分子检测将在法国马赛的Unite des Virus Emergency进行开发，并在加拿大安大略温尼伯的BSL-4机构进行比较和实验评价。最终目标是提出一种用于检测和表征武器化沙粒病毒的即用型诊断试剂盒。

DESCRIPTION (provided by applicant): The objective of the proposed research is to design, develop and evaluate molecular assays to permit rapid and easy detection by real-time RT-PCR of weaponized arenaviruses (AVs) used in a bioterrorism context. Weaponizable AVs include 5 viruses classified in the Category A Pathogen List as defined by the CDC (BSL-4 agents), and all potentially engineered AVs consisting of mutated and/or recombinant and/or reassortant strains. Little genomic data is available for AVs, nevertheless, genetic recombination and reassortment have been observed in natural or experimentally produced AVs. Therefore, a knowledge of the complete genomic sequence of all AVs is the cornerstone of the detection of weaponized AVs. To distinguish BSL-4 AVs from other AVs, of which at least four are naturally circulating is North America and potentially infecting humans, and to detect chimeric engineered AVs it is critical to determine the complete sequences of BSL-4 AVs, but also BSL-2/3 AVs. The realization of this project requires an extensive experience in genomics of RNA viruses, expertise in the design / evaluation process of diagnostic assays of human pathogens, a perfect knowledge of arenavirus genetics, an undisputable working experience of BSL-4 pathogens, and the immediate availability of a BSL-4 laboratory. The molecular assays will be developed at the Unite des Virus Emergents in Marseilles, France and evaluated comparatively and experimentally in the BSL-4 facilities in Winnipeg, Ontario, Canada. The ultimate goal is to propose a ready-to-use diagnostic kit for the detection and characterization of weaponized arenaviruses.