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

肝吸虫、维韦里尼后睾吸虫和华支睾吸虫的护理点核酸检测

• 批准号: 10022036
• 负责人: HyunDae Cho
• 金额: $ 30万
• 依托单位: CROSSLIFE TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-16 至 2020-06-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10022036
• 关键词: Appearance; Base Sequence; Blood; Chemicals; Clonorchis sinensis; Collaborations; Color; DNA; DNA purification; Detection; Development; Devices; Diagnostic; Eye; FDA approved; Fasciola hepatica; Health Personnel; Lateral; Ligation; Molecular; Nucleic Acid Amplification Tests; Nucleic Acids; Opisthorchis viverrini; Paper; Phase; RNA; Reaction; Reader; Recommendation; System; Technology; Telephone; Testing; Training; Validation; base; design; expedited review; experimental study; nanoGold; point of care; stool sample

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.

我们建议开发一种高灵敏度、简单的纸条分子核酸检测方法，使用一种称为TARAamp（模板辅助快速扩增）的平台技术，在医疗点检测肝吸虫（Opisthorchis viverrini）和华支睾吸虫（clon支睾吸虫）。第二阶段开发的最终产品是fda批准的基于核酸的诊断设备纸试纸测试，它包含一个多重探针，可以直接从clbuffer稳定的血液中扩增和检测来自蛇肺和华支睾吸虫的核酸序列，而无需提取RNA。使用TARAamp技术扩增目标序列，该技术由高周转率的模板依赖性化学连接反应、金纳米颗粒（GNP）检测和横向流量读数组成。该测试是快速的，结果是彩色带的外观，可通过眼睛或手机阅读器检测到。该测试可以检测到LOD < 1,000拷贝DNA每次测试。