A New Filter Paper Technology for Flavivirus Collection, Shipping, and Analysis

摘要 几个世纪以来，黄病毒，特别是登革热、寨卡病毒和黄热病一直是一个主要的公共卫生问题 在半热带气候中，包括美国南部的部分地区。目前，这些病毒导致了更多 每年有超过3亿人感染。全球旅行，改变潜在蚊子媒介的地理位置 范围、载体变化导致RNA基因组突变的可能性和新的环境选择性 压力似乎都影响了黄病毒感染向新人类群体的传播。更多 最近，寨卡病毒，被认为与小头畸形症和其他神经出生缺陷有关，已经进入了 强调迫切需要对易感人群进行经济的、国际规模的筛查。 我们认为，迫切需要开发新的技术来收集、运输和回收血液或血液。 含有这种普遍存在的不稳定包膜RNA病毒的产品，在完全不存在任何形式的 通过冷藏进行保护。这项新技术的目标是恢复完整的病毒基因组以支持 病毒载量和核酸检测：为了提高对黄病毒感染的诊断，再加上新的，现在- 廉价的下一代测序方法，为发现病毒基因组如何 可能在选择性压力下发生突变。 这种新的样本收集和保存技术的首要要求是它必须是健壮的 足以保存甚至最不稳定的黄病毒RNA，但又足够便宜，足以支持这两种病毒 人口规模、公共健康筛查和复杂的分子流行病学，以检测突变漂移(通过 下一代测序)，同时以便于在偏远、热带、贫穷 在完全没有低温保存的情况下，易感种群。 GenTegra与Ahlstrom(新生儿筛查医用级滤纸的世界领先者)合作 最近共同开发了一种医用级纤维素纸，它支持DNA在 常温下的血液。在此第二阶段SBIR中，GenTegra将利用与 Ahlstrom，加上我们在开发和实施新的稳定化学物质方面的专业知识， 开发一种全新的滤纸基产品，用于黄病毒RNA的现场采集和保存。至 为了实现这些极具创新性和重大意义的目标，我们提出了三个具体目标，将在两年内执行。 在成功完成后，该计划将生产出可用于生产的产品原型 规模化和国际化商业部署。高校教师队伍的调查与研究环境 合作机构(GenTegra、ATCC和Ahlstrom)在交付方面具有国际水平 这种(21世纪)纸质生物样本收集方法。应该注意的是，虽然这个SBIR是 纯粹专注于黄病毒，该技术可能普遍适用于环境温度收集和 所有的RNA病毒在动物和人类体内的传播。

Abstract The Flaviviruses, especially Dengue, Zika and Yellow Fever have been a major public health concern for centuries in semi and tropical climates, including parts of the southern US. Presently, these viruses are the cause of more than 300 million infections per year. Global travel, alteration of the underlying mosquito vector’s geographical range, the possibility of RNA genome mutation resulting from vector changes and new environmental selective pressures all appear to have influenced the spread of Flavivirus infection to new human populations. More recently, Zika, with the proposed link to microcephaly and other neurological birth defects, has thrust into the spotlight an urgent need for an economical, international-scale screening of susceptible populations. We believe there is a critical need to develop new technology to collect, transport and recover blood or blood products containing this pervasive class of unstable enveloped RNA virus, in the complete absence of any sort of protection via refrigeration. The goal of this new technology is the recovery of intact viral genomes to support viral load and nucleic acid testing: to improve the diagnoses of Flavivirus infection and, coupled with new, now- inexpensive methods of Next Gen sequencing, to provide the real possibility of discovering how the viral genome may be mutating under selective pressure. The overarching requirement for such new sample collection and preservation technology is that it must be robust enough to preserve even the most unstable of the Flavivirus RNAs, yet inexpensive enough to support both population scale public health screening and sophisticated molecular epidemiology to detect mutational drift (via Next-Gen sequencing) while being deployable in a way that facilitates implementation in isolated, tropical, poor, susceptible populations in the complete absence of cryogenic preservation. GenTegra, in collaboration with Ahlstrom (the world-leader in medical grade filter paper for newborn screening) have recently co-developed a medical grade cellulose paper that supports long term stabilization of the DNA in blood at ambient temperature. In this Phase II SBIR, GenTegra will leverage that ongoing collaboration with Ahlstrom, coupled with our expertise in the development and implementation of the new stabilization chemistries, to develop a completely new filter paper based product for Flavivirus RNA field collection and preservation. To achieve those highly Innovative and Significant goals, we propose 3 Specific Aims, to be executed over 2 years. Upon successful completion, this program will yield a product prototype that will be ready for manufacturing scale-up and international commercial deployment. The Investigators and Research Environment among the collaborating institutions (GenTegra, ATCC, and Ahlstrom) are of international caliber with respect to delivering this (21st century) approach to paper-based biosample collection. It should be noted that, although this SBIR is purely focused on Flavivirus, the technology may be generally applicable to ambient temperature collection and transport of all RNA viruses in animals and man.