Fully Integrated Microfluidic In Vitro Diagnostic (IVD) Detection Technology

Various infectious diseases and emerging infectious diseases caused by pathogenic microorganisms remain a major threat to human health and global public health security today. However, many nucleic acid detection products suffer from drawbacks such as multiple operational steps, long time consumption, high dependence on facilities, equipment and personnel, and the need for centralized sample testing.

To address these issues, the team has overcome several key technical bottlenecks, including on-chip reagent storage and controlled release, alcohol-free nucleic acid extraction, nested isothermal amplification, and CRISPR/Cas-based multi-target co-detection. They have developed a "sample in-result out" pathogen nucleic acid microfluidic detection system, making it possible to eliminate cumbersome manual operations in traditional nucleic acid testing, avoid opening lids for amplified products, and reduce reliance on equipment and personnel. The same technical concept has also been applied to the development of fully integrated detection technologies for blood biochemistry and immunology.

Microfluidic Multi-Index Nucleic Acid Detection Technology

Hereditary hearing loss involves numerous hot-spot mutations. Existing methods such as mass spectrometry and sequencing, which rely on multiplex amplification followed by high-throughput analysis, are characterized by complex operational steps, lid-opening procedures for products (prone to false positives), and strict requirements for operators and operating environments. As a result, most hospitals cannot conduct independent testing.

To solve this problem, the applicant has pioneered a microfluidic detection chip for hereditary hearing loss mutations based on centrifugal distribution and thermal sealing blocking. This chip converts multiplex reactions into single-plex reactions, breaking the upper limit of detectable targets in closed chambers through numerous non-interfering microcavities on the chip and addressing the technical limitation of insufficient target coverage in existing technologies. It completely eliminates the exposure of amplified products through physical isolation and, when combined with automated instruments, resolves the high requirements for operators and environments in existing technologies. Hospitals of all sizes can independently carry out testing using this technology, removing barriers to the widespread implementation of hereditary hearing loss gene screening.

The translated and approved detection system, consisting of the "Twenty-Three Genetic Testing Kits for Hereditary Hearing Loss (Microfluidic Chip Method)" and supporting software and hardware, is the world’s first approved microfluidic detection system for hereditary hearing loss genes. It is of great significance for birth defect prevention and control, improving the technical system for tertiary prevention of hereditary hearing loss, and is expected to significantly increase the grassroots penetration rate of hereditary hearing loss gene screening.

It has been adopted by more than 50 clinical institutions nationwide. Governments in Beijing, Chengdu, Shenzhen and other cities have taken the lead in using this system for newborn hereditary hearing loss gene screening, testing nearly all (98-99%) newborns in Beijing since 2022 and making outstanding contributions to safeguarding the quality of the birth population.

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2. Li Zhang, Huili Wang, Sheng Yang, Jiajia Liu, Jie Li, Ying Lu, Jing Cheng*, Youchun Xu*. High-throughput and integrated CRISPR- Cas12a-based molecular diagnosis using a deep learning enabled microfluidic system, ACS Nano, 2024, 18(35):24236-24251.

3. Jiajia Liu, Nan Li, Li Zhang, Ying Lu, Minjie Shen, Yuanyue Zhang, Li Feng, Juhui Jing, Jing Cheng*, Youchun Xu*. A wax interface-enabled one-pot multiplexed nucleic acid testing platform for rapid and sensitive detection of viruses and variants, Small Methods, 2024, 8(8):e2400030.

4. Jiajia Liu, Huili Wang, Li Zhang, Ying Lu, Xu Wang, Minjie Shen, Nan Li, Li Feng, Juhui Jing, Bin Cao, Xiaohui Zou, Jing Cheng*, Youchun Xu*. Sensitive and rapid diagnosis of respiratory virus coinfection using a microfluidic chip-powered CRISPR/Cas12a system, Small, 2022, 18(26):e2200854.

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