This article has been reviewed according to Science X's editorial process and policies. Editors have highlighted the following attributes while ensuring the content's credibility: A research team from The Hong Kong University of Science and Technology (HKUST) has developed an innovative "one-pot" testing platform, known as TEMPO, that could transform highly sensitive nucleic acid testing from a laboratory-based procedure into a simple, single-step test that can be performed at home. With a single reaction tube, users can obtain results comparable to those of professional laboratory tests in as little as 30 minutes.
The breakthrough has the potential to bridge the technological gap between existing rapid tests and laboratory-based nucleic acid diagnostics, offering a more convenient solution for future infectious disease surveillance and genetic screening. TEMPO, short for "Thermodynamically Encoded Molecular Programming for One-Pot Diagnostics," was jointly developed by a research team led by Hsing I-Ming, a professor in the Department of Chemical and Biological Engineering at HKUST, and researchers from The Chinese University of Hong Kong. The platform has already demonstrated applications in the detection of a range of viral infections, including influenza, COVID-19 and HIV, while also opening new possibilities for the rapid screening of hereditary diseases.
The study, titled "Thermodynamically programmed one-pot CRISPR platform for point-of-care SNP genotyping," is published in the journal Nature Communications. Hsing, who led the research and is also the corresponding author of the study, noted, "Conventional laboratory nucleic acid testing involves multiple steps, including amplifying trace amounts of a pathogen's genetic material, identifying target sequences and generating a readable signal. A 'one-pot' test integrates all of these processes into a single reaction tube.
The greatest challenge lies in ensuring that each step takes place in the correct sequence." To precisely control the sequence of reactions, researchers have previously attempted to adjust reagent concentrations, physically separate individual steps or rely on manual intervention to coordinate the process. However, these approaches often lack robustness and are difficult to scale for widespread use. The team adopted a different strategy by harnessing the principles of thermodynamics to enable reactions to proceed automatically in a predetermined order.
This provides a more stable and user-friendly means of controlling reaction timing, allowing nucleic acid tests that would normally require multiple steps to be performed in a single reaction tube. Hsing said, "Think of it like cooking a complex dish in a single pot. The steps have to happen in the right order.
In one-pot CRISPR tests, the process typically involves two key steps: nucleic acid amplification and CRISPR-based detection. These reactions compete for the same ingredients within a sealed reaction tube: amplification needs to build up the target, while the CRISPR enzyme immediately starts consuming it. Start the second step too early and the whole reaction stalls, making it difficult to achieve both speed and sensitivity in a single-step test." At the heart of TEMPO is a pair of thermodynamically engineered DNA primers.
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