the near future,Professor Liu Guozhen's team from the School of Medicine, The Chinese University of Hong Kong (Shenzhen)CRISPR/CasBiosensing technology has made significant breakthroughs in the field of precision medicineRelated research results——“Glass fiber-interfaced CRISPR/Cas biosensing for diverse biomarker detection”The research paper was published ininternational journalCellsub-journalTrends in Biotechnology.

The research team has successfully developed a universal interface based on glass fiber opticsCRISPR/Castesting platform（g-CURS）To achieve ultra-high sensitivity recognition and portable rapid quantitative detection of low abundance multiple analytes (nucleic acids or proteins, etc.), empowering precision medicine and bedside detection（POCT）.thatg-CURSThe system is a continuation of Professor Liu's research team2024Year inCellsub-journalDeviceReported onCRISPRtesting platformGUSCIsystemTechnological upgrade.g-CURSsystemWe have achieved the detection of multiple target nucleic acids and target proteins on the same platform and using the same reagent system, marking a milestoneCRISPRdetection technologyfrom“Single indicator exclusive probe”Restrictions on Jumping to“A pair of universal reagents for detecting multiple types of targets”The standardized operation process greatly reduces research and application costs, providingCRISPRDiagnostic TechnologyPOCTStandardization, scaling up, and commercialization provide new strategies that can be widely applied in pathogen sensitive detection and rapid screening of infectious diseases, early and accurate diagnosis of cancer and chronic diseases, guidance for precise drug use, efficacy evaluation of new drug development, post recovery monitoring of medical art, and animal health testing.

Technical research background

In medical scenarios such as infectious disease surveillance, chronic disease management, early screening of tumors, and precision medicine, highly sensitive detection of various biomarkers such as nucleic acids and proteins in body fluids is crucial. However, the current mainstream methods includeqPCRTheELISAThere are still clinical application problems such as cumbersome operation, dependence on expensive instruments, large sample demand, long detection time, and unsuitability for resource poor scenarios.

althoughCRISPRTechnology has gradually been applied to in vitro diagnostics, but traditional strategies need to be designed separately for each targetcrRNAAnd activators, for the detection of low abundance test substances, highly rely on nucleic acid pre amplification programs, resulting in complex screening and optimization steps in the early design, long detection time, low application efficiency, and poor universality. For this purpose, Professor Liu's research team has developed aPortable without pre amplificationCRISPR/CasQuick detection platform:g-CURSNot only does it solve the problem of traditional detection methods in clinical applications, but it also expandsCRISPR/CasThe application of biosensing technology in precision medicine enables the simultaneous detection of multiple biomarkers in the same system.

g-CURSTechnical principles and highlights

g-CURSThe detection platform uses glass fiber as a carrier and utilizesCRISPR/Cas12aExponential signal amplification technology, utilizing the same setcrRNAandssDNAActivating the sub reagent system can detect different nucleic acids and multiple protein targets without the need for personalized design for different targetsCRISPR RNAScreen for corresponding target sitesChoose;The chip channel structure within the detection platform is constructed by Mofang Precision Surface Projection MicroStereoscopic Lithography (P μ SL) Technology (nanoArch)^®S140， Precision: 10 μ m) After preparing the mold, it is flipped over by PDMSThe nucleic acid recognition process adoptsT4 DNALigase for precise identification of single nucleotide differences. No need for pre amplification,CRISPR/Cas利用AutoCARThe principle of cascade amplification andCONANReaction network induces signal exponential amplification loop, achieving30Rapid isothermal amplification within minutes; Constructing capture antibodies and labeling on the surface of glass fiber optic cablesssDNAThe detection of antibodies“Immune sandwich structure”, actuallynowProtein signal conversion and amplification; Using self-made methodsPortable fluorescence analyzer reads the signal of the test object, equipped with a high-resolution fixed focus lens and wireless module, and paired with end-user mobile phonesAppRealize result visualization and intelligent diagnosis, support independent operation by non professionals, and meet the requirementsPOCTapplicationScenario requirements.

clinical applicationperformance evaluation

g-CURSCapable of efficient and accurate clinical testing performance, with high specificity recognitionSARS-CoV-2 DeltamutantcDNAandHPV16 E7Gene fragments, testing time is approximately30minute; Accurately identifiable6Mutations within one nucleotide range have the ability to distinguish single nucleotide mismatchesHPV16In clinical samples, the test results are consistent withqPCRagreement. Applied to serum samples of Parkinson's disease patients,g-CURSCan achieve high-sensitivity detection of low abundance inflammatory proteins in the membrane of brain derived exosomes（IL-6TheTNF-αTheIFN-γwait）Support low sample size multi indicator co testing. Compared to commercial useELISACompared to the reagent kit, the detection sensitivity has been improved2-3By an order of magnitude, the detection time has been shortened80%,The required sample volume is greatly reduced (only5ML）The cost of testing has been reduced90%, achieving portabilityCRISPR/CasSimultaneous detection of multiple substances using in vitro diagnostic technology. This breakthroughCRISPR/CasThe detection platform is expected to promote precision medicine from the laboratory to the community, providing more convenient and economical solutions for early diagnosis and dynamic monitoring of diseases.

The author of this paper is from Hong KongChinese majorPhD student majoring in Biological Sciences at Shenzhen Medical CollegeYu JianThe corresponding author of the paper is the School of Medicine, The Chinese University of Hong Kong (Shenzhen)Professor Liu GuozhenThis work has received funding from the National Natural Science Foundation of China, the Guangdong Provincial Basic and Applied Basic Research Fund Enterprise Joint Fund, and the Guangdong Provincial Government“The Pearl River Talent Plan”Shenzhen Bay Laboratory Open Fund, Hong Kong Zhongshen-Funding from the Boya Life Joint Laboratory Fund and the University Development Fund.