Point of Care Nucleic Acids Test for the Liver Flukes Opisthorchis viverrini and Clonorchis

我们建议开发一种高灵敏度、简单的试纸条分子核酸检测方法，使用称为TARAamp（模板辅助快速扩增）的平台技术在护理点检测肝吸虫后睾吸虫和华支睾吸虫。第2阶段开发的最终产品是FDA批准的基于核酸的诊断设备试纸条检测，其含有多重探针，可以直接从CL缓冲液稳定的血液中扩增和检测来自猫背睾吸虫和中华克隆虫的核酸序列，而无需RNA提取。使用TARAamp技术扩增靶序列，该技术由具有高周转率的模板依赖性化学连接反应、金纳米颗粒（GNP）检测和侧向流读出组成。该测试是快速的，并导致出现可通过眼睛或手机阅读器检测到的彩色条带。该测试可以检测LOD <1，000份DNA。 在第二阶段，我们将进行大规模的验证实验，以获得世界卫生组织对我们产品的快速审查批准和采购建议。在第2阶段，我们将为我们开发的检测卡开发CrossLife移动的阅读器（与NovarumDx合作），通过消除任何主观性，为用户提供明确的结果。我们简单易用的技术可以补充测试编排，这有助于进一步降低系统中的错误率，对于培训较少的医疗工作者尤其重要。

We propose the development of a highly sensitive, simple paper strip molecular nucleic acid test that detects the liver fluke Opisthorchis viverrini and Clonorchis sinensis at the point-of-care using a platform technology called TARAamp (Template-Assisted Rapid Amplification). The final product to be developed in Phase 2 is an FDA-approved nucleic acid-based diagnostic device paper dipstick test that contains a multiplex of probe that can amplify and detect nucleic acid sequences from the Opisthorchis viverrini and Clonorchis sinensis directly from CLBuffer-stabilized blood without RNA extraction. The target sequences are amplified using the TARAamp technology which is composed of a template-dependent chemical ligation reaction with high turnover, and gold nanoparticles (GNP) detection and lateral flow readout. The test is rapid, and results in the appearance of colored bands that are detectable by eye or phone Reader. The test can detect LOD is < 1,000 copies of DNA per test. In Phase 2, we will conduct a large-scale validation experiment to obtain expedited review approval and recommendation for procurement of our product from the WHO. In Phase 2, we will develop a CrossLife mobile reader (collaboration with NovarumDx) for our developed test cassette that will provide the user with unambiguous results, by removing any subjectivity. Our simple to use technology may be supplemented with test choreography, that aids to reduce error rates in the system even further, and is particularly important for less-trained healthcare workers.