Direct molecular detection of DNA for blood genotyping

仅在魁北克省，每年就有超过25万单位的血液被输入。为了向医院提供安全的血液，血库设施必须检测每一位献血者和受血者是否存在几种红细胞抗原。由于人口老龄化、输血次数的增加以及用于测试这些抗原的现有技术的复杂性和有限的样品通量，匹配血液的采购代表了血库日益严重的操作问题。本文提出的项目的目的是开发一种快速简单的方法，用于使用我们的研究团队最近证明的称为荧光链式反应（FCR）的分子传感方案来鉴定血液基因型。这种新的检测方案结合了特定的寡核苷酸探针和光学聚合物传感器，并能够在5分钟内检测少至5个拷贝的遗传物质，具有区分单核苷酸多态性所需的特异性。如检测能力，这是几个数量级领先于目前正在开发的任何其他方法，有可能提供快速，可靠和负担得起的分子分型程序的浓度水平，使事先的化学扩增或标记将不再是必要的。该研究项目将专注于增加FCR对实验条件变化的特异性和稳健性，以及代表献血者和受血者临床相关抗原基因分型中发现的样品组成。通过本提案中所述的研究和实验继续发展这项技术将对生物技术/生物医学部门和公共卫生问题产生巨大影响，我们相信，包括我们的合作伙伴Hema-Quebec在内的各个领域的科学家和工业将从这项技术中受益匪浅。

More than 250,000 blood units are transfused each year in the province of Quebec alone. In order to provide safe blood to hospitals, blood bank facilities must test every donor and recipient for the presence of several red cell antigens. Because of population aging, of the increasing number of transfusions and of the complexity and limited sample throughput of currently available technology used to test for these antigens, the procurement of matched blood represents a growing operational problem for blood banks. The purpose of the project proposed herein is to develop a rapid and simple method for the identification of blood genotypes using a molecular sensing scheme called Fluorescence Chain Reaction (FCR) recently demonstrated by our research team. This novel detection scheme combines specific oligonucleotide probes and an optical polymeric transducer and is able to detect as little as 5 copies of genetic material in 5 minutes with the specificity required to distinguish single nucleotide polymorphisms. Such as detection capability, which is several orders of magnitude ahead of any other approach currently in development, has the potential to provide fast, reliable and affordable molecular typing procedures at concentration levels such that prior chemical amplification or tagging will no longer be necessary. This research project will focus on increasing FCR's specificity and robustness towards variations in experimental conditions and for sample compositions that are representative of those found in the genotyping of clinically relevant antigens of blood donors and recipients. Continued development of this technology through the studies and experiments described in the present proposal will have a massive impact in the biotechnology/biomedical sectors and on public health issues, and we believe that scientists and industries in various fields, including our partner Hema-Quebec, will greatly benefit from this technology.