MRIGlobal, 65 West Watkins Mill Road, Gaithersburg, MD, 20878, USA.
MRIGlobal, 1470 Treeland Blvd. SE, Palm Bay, FL, 32909, USA.
Sci Rep. 2018 Apr 3;8(1):5417. doi: 10.1038/s41598-018-23641-7.
The future of infectious disease surveillance and outbreak response is trending towards smaller hand-held solutions for point-of-need pathogen detection. Here, samples of Culex cedecei mosquitoes collected in Southern Florida, USA were tested for Venezuelan Equine Encephalitis Virus (VEEV), a previously-weaponized arthropod-borne RNA-virus capable of causing acute and fatal encephalitis in animal and human hosts. A single 20-mosquito pool tested positive for VEEV by quantitative reverse transcription polymerase chain reaction (RT-qPCR) on the Biomeme two3. The virus-positive sample was subjected to unbiased metatranscriptome sequencing on the Oxford Nanopore MinION and shown to contain Everglades Virus (EVEV), an alphavirus in the VEEV serocomplex. Our results demonstrate, for the first time, the use of unbiased sequence-based detection and subtyping of a high-consequence biothreat pathogen directly from an environmental sample using field-forward protocols. The development and validation of methods designed for field-based diagnostic metagenomics and pathogen discovery, such as those suitable for use in mobile "pocket laboratories", will address a growing demand for public health teams to carry out their mission where it is most urgent: at the point-of-need.
传染病监测和疫情应对的未来趋势是针对现场病原体检测，使用更小的手持式解决方案。这里，对采集自美国佛罗里达州南部的库蚊进行了检测，以确定是否存在委内瑞拉马脑炎病毒（VEEV）。VEEV 是一种先前被武器化的节肢动物传播的 RNA 病毒，能够导致动物和人类宿主急性和致命的脑炎。在 Biomeme two3 上，通过定量逆转录聚合酶链反应（RT-qPCR）对 20 只蚊子的单一混合样本进行检测，结果显示 VEEV 呈阳性。对病毒阳性样本进行无偏倚的宏转录组测序，结果显示该样本中含有伊蚊病毒（EVEV），这是一种 VEEV 血清群中的甲病毒。我们的结果首次证明，使用基于序列的无偏检测方法，以及现场向前协议，可直接从环境样本中对高后果生物威胁病原体进行检测和亚型分析。开发和验证适合现场诊断宏基因组学和病原体发现的方法，例如适合在移动“口袋实验室”中使用的方法，将满足公共卫生团队在最需要的地方开展工作的需求：现场。
