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

Crosslife Technologies, Inc., Carlsbad CA

Investigators

Abstract

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.