Recent advances in CRISPR- and RCA-based biosensing chips and devices for POCT and in situ detection

doi:10.1007/s44275-025-00038-z

Moore and More ›› 2026, Vol. 2 ›› Issue (1): 84-114.DOI: 10.1007/s44275-025-00038-z

Recent advances in CRISPR- and RCA-based biosensing chips and devices for POCT and in situ detection

Yuming Huang¹^,³, Tianyi Yin², Yuhao Wen¹^,³, Shenye Qian¹^,³, Yuanjie Wu¹^,³, Xingkai Hao¹^,³, Wen Chen¹^,³, Bing Li⁴, Zhengdong Li⁵^,^*(), Sami Ramadan⁶^,^*(), Lizhou Xu¹^,³^,^*()

¹ ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University , Hangzhou 311200, China
² State Key Laboratory of Complex, Severe, and Rare Diseases, Biomedical Engineering Facility of National Infrastructures for Translational Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100730, China
³ College of Biosystems Engineering and Food Science, Zhejiang University , Hangzhou 310058, China
⁴ Department of Chemistry, Institute for Materials Discovery, University College London , London WC1E 7JE, UK
⁵ Precision Medical Center, Guangyuan Central Hospital , Guangyuan 628000, China
⁶ Department of Materials, Imperial College London , London SW7 2AZ, UK

Received:2025-05-27 Revised:2025-09-15 Accepted:2025-09-24 Published:2026-03-20 Online:2026-05-12
Contact:
^* Zhengdong Li (Zhengdongli5193@163.com)
^* Sami Ramadan (s.ramadan@imperial.ac.uk)
^* Lizhou Xu (lzxu@zju.edu.cn)
About author:
Yuming Huang is a student majoring in Bioengineering at Zhejiang University. His current research interests include the development of advanced biosensing tools for POCT and in situ analysis.
Tianyi Yin is a postdoctoral fellow at State Key Laboratory of Complex, Severe, and Rare Diseases, Peking Union Medical College Hospital. She obtained her Ph.D. degree in Materials Science and Engineering in 2024 from Imperial College London. She was awarded the Constance Fligg Tipper Centenary Memorial Prize for her postgraduate research. Her current research focuses on the design, development, and biomedical applications of biosensors.
Yuhao Wen obtained his M.S. degree in Materials and Chemicals from the Harbin Institute of Technology in 2023. In 2024, he joined the Hangzhou Global Scientific and Technological Innovation Center in Zhejiang University. His current research interest is focused on the construction and application of biochemical sensors and related microdevices.
Shenye Qian is a master’s student at Zhejiang University. He obtained his B.S. degree in Food Quality and Safety from Soochow University in 2024. He is currently pursuing a master’s degree in Food Engineering at the ZJU-Hangzhou Global Scientific and Technological Innovation Center. His recent research focuses on the design, development, and application of biosensors.
Yuanjie Wu graduated from the Zhejiang Chinese Medical University with a master’s degree in Basic Medicine in 2025. His main research focuses on bio-detection, drug delivery, and pharmacology. In 2025, he joined the Hangzhou International Science and Technology Innovation Center of Zhejiang University to engage in drug delivery and biological detectionrelated work.
Xingkai Hao is currently a postdoctoral researcher at Zhejiang University, focusing on highsensitivity biosensing technologies for applications in food safety and medical diagnostics. He obtained his Ph.D. degree in Chemical Engineering from the University of Ottawa in 2023. His recent research involves developing rapid detection systems for pathogens and other contaminants in food and beverages, with a particular emphasis on improving the quality control of dairy products.
Wen Chen is currently a postdoctoral researcher at Zhejiang University, focused on extracting active ingredients in functional foods and evaluating their efficacy, as well as developing highsensitivity biosensing technologies for medical diagnostics. She obtained her Ph.D. degree in Food Science from Zhejiang University in 2023. Her recent research has concentrated on developing rapid detection systems for miRNA biomarkers to enhance the quality control of dairy products and on exosomes for targeted bioactive ingredient delivery.
Bing Li is a full-time lecturer in advanced biosensing technology at UCL. He received his Ph.D. degree from University of Plymouth, UK, in 2016. Then, he moved to Imperial College London as a postdoctoral research associate (PDRA) to develop smart biosensing systems for surgical robotics and medical tools. In 2019, he secured a prestigious independent Edmond and Lily Safra Research Fellowship at Imperial College London to develop the next-generation biosensing diagnostics for the detection of dementia biomarkers. His group has been focusing on the development of various point-of-care/wearable/ implantable sensing systems for disease monitoring and other biomedical applications. He also holds an Emerging Leader position with the UK Dementia Research Institute and an Honorary Lecturer with the Department of Brain Sciences at Imperial College London.
Zhengdong Li is currently a research director at the Precision Medical Center, Guangyuan Central Hospital. He completed his medicine study at the Faculty of Medicine, University of Rostock. He holds a Doctor of Biochemistry from Freie Universität Berlin, Germany. Afterwards, he completed his postdoctoral research at the Helmholtz Center for Biomaterials and the Brandenburg Center for Regenerative Medicine in Berlin, Germany. His research interests include molecular diagnosis, precision medicine, and biomaterials in biomedical areas.
Sami Ramandan is currently a senior research associate at Department of Materials, Faculty of Engineering, Imperial College London. His research focus is on developing innovative biosensors for early disease detection and diagnosis. His research utilizes two-dimensional (2D) nanomaterials to create ultrasensitive, cost-effective, and user-friendly biosensor technologies that can be translated into point-of-care diagnosis.
Lizhou Xu is currently a professor at Hangzhou Global Scientific and Technological Innovation Center & College of Biosystems Engineering and Food Science, Zhejiang University. He was a research associate at Department of Materials, Imperial College London during 2020–2021 studying novel materials and new biosensors. He worked as a KC Wong Fellow (2016–2017) and a Marie Skłodowska-Curie Fellow (2017–2019) at the School of Cancer & Pharmaceutical Sciences, King’s College London. His current research group focuses on the advanced biosensing technologies and biochips for high performance detection in food, agriculture, and medicine.

Abstract

Abstract:

The synergy of clustered regularly interspaced short palindromic repeats (CRISPR) and rolling circle amplification (RCA) has been considered the cutting edge of molecular diagnostics. These biosensing assays, integrated with other techniques, are widely used in various biosensors for the detection of multiple targets, including nucleic and non-nucleic acid targets. Some biosensing platforms have demonstrated detection performance comparable to that of the gold standard techniques, showing the potential for application in the field of point-of-care testing (POCT). Herein, we review the recent advances over the past 5 years in CRISPR- and RCA-based biosensing chips and devices, with a focus on their implementation in POCT and in situ applications. We classify these systems based on target type, compare their performance metrics, and discuss key technological features. Based on relevant commercial products, we also outline the current challenges that these biosensors may face during commercialization and propose some potential solutions and future research directions.

Key words: POCT, In situ detection, Biosensing chips and devices, CRISPR, RCA

Yuming Huang, Tianyi Yin, Yuhao Wen, Shenye Qian, Yuanjie Wu, Xingkai Hao, Wen Chen, Bing Li, Zhengdong Li, Sami Ramadan, Lizhou Xu. Recent advances in CRISPR- and RCA-based biosensing chips and devices for POCT and in situ detection[J]. Moore and More, 2026, 2(1): 84-114.

Yuming Huang, Tianyi Yin, Yuhao Wen, Shenye Qian, Yuanjie Wu, Xingkai Hao, Wen Chen, Bing Li, Zhengdong Li, Sami Ramadan, Lizhou Xu. [J]. Moore and More, 2026, 2(1): 84-114.

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References 217

[1]	Dong H , Lei J , Ding L , Wen Y , Ju H , Zhang X (2013) MicroRNA: function, detection, and bioanalysis. Chem Rev 113: 6207-6233. https://doi.org/10.1021/cr300362f
[2]	Mitra P , Sharma P (2021) POCT in developing countries. Electron J Int Fed Clin Chem Lab Med 32(2): 195-199
[3]	Kumar A , Parihar A , Panda U , Parihar DS (2022) Microfluidics-based point-of-care testing (POCT) devices in dealing with waves of COVID-19 pandemic: the emerging solution. ACS Appl Bio Mater 5: 2046-2068. https://doi.org/10.1021/acsabm.1c01320
[4]	Strohmaier-Nguyen D , Horn C , Baeumner AJ (2025) Innovations in one-step point-of-care testing within microfluidics and lateral flow assays for shaping the future of healthcare. Biosens Bioelectron 267: 116795. https://doi.org/10.1016/j.bios.2024.116795
[5]	Pokhrel P , Hu C , Mao H (2020) Detecting the coronavirus (COVID-19). ACS Sens 5: 2283-2296. https://doi.org/10.1021/acssensors.0c01153
[6]	Yan T , Zhang G , Chai H , Qu L , Zhang X (2021) Flexible biosensors based on colorimetry, fluorescence, and electrochemistry for point-of-care testing. Front Bioeng Biotechnol 9: 753692. https://doi.org/10.3389/fbioe.2021.753692
[7]	Sachdeva S , Davis RW , Saha AK (2021) Microfluidic point-of-care testing: commercial landscape and future directions. Front Bioeng Biotechnol 8: 602659. https://doi.org/10.3389/fbioe.2020.602659
[8]	Lee S , Bi L , Chen H , Lin D , Mei R , Wu Y et al (2023) Recent advances in point-of-care testing of COVID-19. Chem Soc Rev 52: 8500-8530. https://doi.org/10.1039/d3cs00709j
[9]	Dang VA , Vu Khanh Q , Nguyen V-H , Nguyen T , Nguyen DC (2023) Intelligent healthcare: integration of emerging technologies and internet of things for humanity. Sensors (Basel) 23: 4200. https://doi.org/10.3390/s23094200
[10]	Jain S , Nehra M , Kumar R , Dilbaghi N , Hu T , Kumar S et al (2021) Internet of medical things (IoMT)-integrated biosensors for point-of-care testing of infectious diseases. Biosens Bioelectron 179: 113074. https://doi.org/10.1016/j.bios.2021.113074
[11]	Chakraborty T , Roy P , Nesa N (2023) Iomt-based biochip-integrated point-of-care testing for smart healthcare. IEEE Internet Things J 10: 22740-22747. https://doi.org/10.1109/JIOT.2023.3304702
[12]	Haase AT , Retzel EF , Staskus KA (1990) Amplification and detection of lentiviral DNA inside cells. Proc Natl Acad Sci U S A 87: 4971-4975. https://doi.org/10.1073/pnas.87.13.4971
[13]	Tedesco I , Zito Marino F , Ronchi A , Duarte Neto AN , Dolhnikoff M , Municinò M et al (2023) COVID-19: detection methods in post-mortem samples. Pathologica 115: 263-274. https://doi.org/10.32074/1591-951X-933
[14]	Schaefer I-M , Padera RF , Solomon IH , Kanjilal S , Hammer MM , Hornick JL et al (2020) In situ detection of SARS-CoV-2 in lungs and airways of patients with COVID-19. Mod Pathol 33: 2104-2114. https://doi.org/10.1038/s41379-020-0595-z
[15]	Jensen E (2014) Technical review: in situ hybridization. Anat Rec Hoboken 297: 1349-1353. https://doi.org/10.1002/ar.22944
[16]	Weng Z , You Z , Yang J , Mohammad N , Lin M , Wei Q et al (2023) CRISPR-cas biochemistry and CRISPR-based molecular diagnostics. Angew Chem Int Ed Engl 62: e202214987. https://doi.org/10.1002/anie.202214987
[17]	Pacesa M , Pelea O , Jinek M (2024) Past, present, and future of CRISPR genome editing technologies. Cell 187: 1076-1100. https://doi.org/10.1016/j.cell.2024.01.042
[18]	Jinek M , Chylinski K , Fonfara I , Hauer M , Doudna JA , Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337: 816-821. https://doi.org/10.1126/science.1225829
[19]	Kaminski MM , Abudayyeh OO , Gootenberg JS , Zhang F , Collins JJ (2021) CRISPR-based diagnostics. Nat Biomed Eng 5: 643-656. https://doi.org/10.1038/s41551-021-00760-7
[20]	Liu J , Xie G , Lv S , Xiong Q , Xu H (2023) Recent applications of rolling circle amplification in biosensors and DNA nanotechnology. TrAC Trends Anal Chem 160: 116953. https://doi.org/10.1016/j.trac.2023.116953
[21]	Hansen MF , Dufva M , Fock J , Minero GAS , Tian B (2020) CRISPR-Cas12a based internal negative control for nonspecific products of exponential rolling circle amplification. Nucleic Acids Res 48: e30. https://doi.org/10.1093/nar/gkaa017
[22]	Gupta N , Augustine S , Narayan T , O’Riordan A , Das A , Kumar D et al (2021) Point-of-care PCR assays for COVID-19 detection. Biosensors 11: 141. https://doi.org/10.3390/bios11050141
[23]	Zhao Y , Wu W , Tang X , Zhang Q , Mao J , Yu L et al (2023) A universal CRISPR/Cas12a-powered intelligent point-of-care testing platform for multiple small molecules in the healthcare, environment, and food. Biosens Bioelectron 225: 115102. https://doi.org/10.1016/j.bios.2023.115102
[24]	Xu L , Duan J , Chen J , Ding S , Cheng W (2021) Recent advances in rolling circle amplification-based biosensing strategies – a review. Anal Chim Acta 1148: 238187. https://doi.org/10.1016/j.aca.2020.12.062
[25]	Long Y , Tao S , Shi D , Jiang X , Yu T , Long Y et al (2024) Special RCA based sensitive point-of-care detection of HPV mRNA for cervical cancer screening. Aggregate 5(4): e569. https://doi.org/10.1002/agt2.569
[26]	Deng Z , Liao W , Bu L , Li J , Li J , Wang L et al (2024) Advancements in CRISPR-diagnostic techniques for rapid on-site monitoring of environmental virus. TrAC Trends Anal Chem 181: 118046. https://doi.org/10.1016/j.trac.2024.118046
[27]	Dalton J (2021) Communications with lab and POCT users. Pract Lab Med 25: e00223. https://doi.org/10.1016/j.plabm.2021.e00223
[28]	Shaw JLV (2015) Practical challenges related to point of care testing. Pract Lab Med 4: 22-29. https://doi.org/10.1016/j.plabm.2015.12.002
[29]	Guo L , Zhao Y , Huang Q , Huang J , Tao Y , Chen J et al (2024) Electrochemical protein biosensors for disease marker detection: progress and opportunities. Microsyst Nanoeng 10: 65. https://doi.org/10.1038/s41378-024-00700-w
[30]	Yammouri G , Ait Lahcen A (2024) AI-reinforced wearable sensors and intelligent point-of-care tests. J Pers Med 14: 1088. https://doi.org/10.3390/jpm14111088
[31]	Zhu Z , Yang L (2024) Recent progress in molecular diagnostics: the synergy of rolling circle amplification and CRISPR/Cas systems (2018–2024) – a concise review. TrAC Trends Anal Chem 180: 117902. https://doi.org/10.1016/j.trac.2024.117902
[32]	Zeng F , Nijiati S , Tang L , Ye J , Zhou Z , Chen X (2023) Ferroptosis detection: from approaches to applications. Angew Chem Int Ed Engl 62: e202300379. https://doi.org/10.1002/anie.202300379
[33]	Wang Z-Y , Li D-L , Tian X , Li Y , Zhang C-Y (2022) Single-molecule counting of FTO in human breast tissues based on a rolling circle transcription amplification-driven clustered regularly interspaced short palindromic repeat─Cas12a. Anal Chem 94: 11425-11432. https://doi.org/10.1021/acs.analchem.2c02578
[34]	Khan SF , Rathod P , Gupta VK , Khedekar PB , Chikhale RV (2024) Evolution and impact of nucleic acid amplification test (NAAT) for diagnosis of coronavirus disease. Anal Chem 96: 8124-8146. https://doi.org/10.1021/acs.analchem.3c05225
[35]	Li M , Yin F , Song L , Mao X , Li F , Fan C et al (2021) Nucleic acid tests for clinical translation. Chem Rev 121: 10469-10558. https://doi.org/10.1021/acs.chemrev.1c00241
[36]	Yan H , Wen Y , Tian Z , Hart N , Han S , Hughes SJ et al (2023) A one-pot isothermal Cas12-based assay for the sensitive detection of microRNAs. Nat Biomed Eng 7: 1583-1601. https://doi.org/10.1038/s41551-023-01033-1
[37]	Xu M , Wang R , Li Y (2017) Electrochemical biosensors for rapid detection of Escherichia coli O157:H7 . Talanta 162: 511-522. https://doi.org/10.1016/j.talanta.2016.10.050
[38]	Shi J , Lei C , Fan W , Sun Y , Liu C (2024) Ultrasensitive protein and exosome analysis based on a rolling circle amplification assisted-CRISPR/Cas12a strategy. Talanta 273: 125906. https://doi.org/10.1016/j.talanta.2024.125906
[39]	Suea-Ngam A , Deck L-T , Howes PD , deMello AJ (2020) An ultrasensitive non-noble metal colorimetric assay using starch-iodide complexation for Ochratoxin A detection. Anal Chim Acta 1135: 29-37. https://doi.org/10.1016/j.aca.2020.08.028
[40]	Bruch R , Baaske J , Chatelle C , Weber W , Dincer C , Urban G (2019) Electrochemical biosensor for CRISPR/Cas13a powered miRNA diagnostics. In: 2019 IEEE Sensors, Montreal, QC, Canada, 2019. IEEE, pp 1-3. https://doi.org/10.1109/SENSORS43011.2019.8956561
[41]	Bruch R , Baaske J , Chatelle C , Meirich M , Madlener S , Weber W et al (2019) CRISPR/Cas13a-powered electrochemical microfluidic biosensor for nucleic acid amplification-free miRNA diagnostics. Adv Mater 31: e1905311. https://doi.org/10.1002/adma.201905311
[42]	Zhou Y , Xie S , Liu B , Wang C , Huang Y , Zhang X et al (2023) Chemiluminescence sensor for miRNA-21 detection based on CRISPR-Cas12a and cation exchange reaction. Anal Chem 95: 3332-3339. https://doi.org/10.1021/acs.analchem.2c04484
[43]	Zhao W , Zhang X , Tian R , Li H , Zhong S , Yu R (2023) The sensor platform combined with dual signal amplification and based on UCNPs and CRISPR/Cas12a for miRNA-21 detection. Sens Actuator B-Chem 393: 134238. https://doi.org/10.1016/j.snb.2023.134238
[44]	Liu R , Wang X , Wang S , Xie L , Zhao P , Li L et al (2024) Rolling circle amplification assisted CRISPR/Cas12a dual-cleavage photoelectrochemical biosensor for highly sensitive detection of miRNA-21. Anal Chim Acta 1287: 342125. https://doi.org/10.1016/j.aca.2023.342125
[45]	Liu M , Ma W , Zhou Y , Liu B , Zhang X , Zhang S (2022) A label-free photoelectrochemical biosensor based on CRISPR/Cas12a system responsive deoxyribonucleic acid hydrogel and “click” chemistry. ACS Sens 7: 3153-3160. https://doi.org/10.1021/acssensors.2c01636
[46]	Shen H , Yang H , Qileng A , Ma Y , Liang H , Meng J (2022) Programmable readout sensor for microRNA: CRISPR/Cas12a-assisted multi-amplification strategy activated photoelectrochemistry-colorimetry detection. Sens Actuator B Chem 371: 132585. https://doi.org/10.1016/j.snb.2022.132585
[47]	Li Y , Zeng Z , Lv X , Jiang H , Li A , Liu Y (2025) A POCT assay based on commercial HCG strip for miRNA21 detection by integrating with RCA-HCR cascade amplification and CRISPR/Cas12a. Microchim Acta 192: 73. https://doi.org/10.1007/s00604-024-06922-z
[48]	Dong J , Li X , Zhou S , Liu Y , Deng L , Chen J et al (2023) CRISPR/Cas12a-powered EC/FL dual-mode controlled-release homogeneous biosensor for ultrasensitive and cross-validated detection of messenger ribonucleic acid. Anal Chem 95: 12122-12130. https://doi.org/10.1021/acs.analchem.3c02335
[49]	Wang S , Li H , Dong K , Shu W , Zhang J , Zhang J et al (2023) A universal and specific RNA biosensor via DNA circuit-mediated PAM-independent CRISPR/Cas12a and polyA-rolling circle amplification. Biosens Bioelectron 226: 115139. https://doi.org/10.1016/j.bios.2023.115139
[50]	Ma W , Liu M , Xie S , Liu B , Jiang L , Zhang X et al (2022) CRISPR/Cas12a system responsive DNA hydrogel for label-free detection of non-glucose targets with a portable personal glucose meter. Anal Chim Acta 1231: 340439. https://doi.org/10.1016/j.aca.2022.340439
[51]	Qing M , Chen SL , Sun Z , Fan Y , Luo HQ , Li NB (2021) Universal and programmable rolling circle amplification-CRISPR/Cas12a-mediated immobilization-free electrochemical biosensor. Anal Chem 93: 7499-7507. https://doi.org/10.1021/acs.analchem.1c00805
[52]	Wang H , Hang X , Wang H , Peng J , Yu H , Wang L (2024) Label/immobilization-free Cas12a-based electrochemiluminescence biosensor for sensitive DNA detection. Talanta 275: 126114. https://doi.org/10.1016/j.talanta.2024.126114
[53]	Zhang Y , Hu C , Yin Y , Ren K , He Y , Gao Y et al (2024) CRISPR/Cas12a-responsive smart DNA hydrogel for sensitive electrochemiluminescence detection of the Huanglongbing outer membrane protein gene. Anal Chem 96: 11611-11618. https://doi.org/10.1021/acs.analchem.4c02489
[54]	You J , Park H , Lee H , Jang K , Park J , Na S (2023) Sensitive and selective DNA detecting electrochemical sensor via double cleaving CRISPR Cas12a and dual polymerization on hyperbranched rolling circle amplification. Biosens Bioelectron 224: 115078. https://doi.org/10.1016/j.bios.2023.115078
[55]	Deng L , Zhou S , Dong J , Liu Y , Huang Z , Sun H et al (2023) CRISPR/Cas12a and primer-assisted rolling circle amplification integrated ultra-sensitive dual-signal sensing platform for EGFR 19 detection. Anal Chim Acta 1279: 341755. https://doi.org/10.1016/j.aca.2023.341755
[56]	Guo K-H , Chen P-H , Lin C , Chen C-F , Lee IR , Yeh Y-C (2018) Determination of gold ions in human urine using genetically engineered microorganisms on a paper device. ACS Sens 3: 744-748. https://doi.org/10.1021/acssensors.7b00931
[57]	Xu J , Wang M , Li M , Yang J , Yang L (2023) Paper-based chiral biosensors using enzyme encapsulation in hydrogel network for point-of-care detection of lactate enantiomers. Anal Chim Acta 1279: 341834. https://doi.org/10.1016/j.aca.2023.341834
[58]	Jiang N , Ahmed R , Damayantharan M , Ünal B , Butt H , Yetisen AK (2019) Lateral and vertical flow assays for point-of-care diagnostics. Adv Healthc Mater 8: e1900244. https://doi.org/10.1002/adhm.201900244
[59]	Wang X , Zhao C , Yin N , Wang X , Shu Y , Wang J (2025) Dual miRNAs imaging platform based on HRCA-Cas12a by replacing PAM with bubble to reduce false positive. Anal Chem 97: 3053-3062. https://doi.org/10.1021/acs.analchem.4c06162
[60]	Yang G , Song T , Wang M , Li M , Su Q , Xie Z et al (2022) Recent advancements in nanosystem-based molecular beacons for RNA detection and imaging. ACS Appl Nano Mater 5: 3065-3086. https://doi.org/10.1021/acsanm.1c03966
[61]	Xue Y , Xie H , Wang Y , Feng S , Sun J , Huang J et al (2022) Novel and sensitive electrochemical/fluorescent dual-mode biosensing platform based on the cascaded cyclic amplification of enzyme-free DDSA and functional nucleic acids. Biosens Bioelectron 218: 114762. https://doi.org/10.1016/j.bios.2022.114762
[62]	Dong J , Zhou S , Liu Y , Deng L , Huang Z , Chen J et al (2023) A self-supply crRNA-mediated CRISPR/Cas12a-driven controlled-release homogeneous biosensor for ultrasensitive detection of microRNA. Chem Eng J 471: 144507. https://doi.org/10.1016/j.cej.2023.144507
[63]	Jiang M , Hong X , Gao Y , Kho AT , Tantisira KG , Li J (2024) Pirna associates with immune diseases. Cell Commun Signal 22: 347. https://doi.org/10.1186/s12964-024-01724-5
[64]	Tian Z , Luo J , Zhang C , Li Y , Hu S , Li Y (2024) Photonic crystal-enhanced fluorescence biosensor with logic gate operation based on one-pot cascade amplification DNA circuit for enzyme-free and ultrasensitive analysis of two microRNAs. Talanta 277: 126428. https://doi.org/10.1016/j.talanta.2024.126428
[65]	Deng Y , Zhou T , Hu K , Peng Y , Jia X , Yang J et al (2024) An electrochemical biosensor designed with entropy-driven autocatalytic DNA circuits for sensitive detection of ovarian cancer-derived exosomes. Biosens Bioelectron 250: 116060. https://doi.org/10.1016/j.bios.2024.116060
[66]	Liu R , Han H , Liu F , Lv Z , Wu K , Liu Y et al (2020) Positive rate of RT-PCR detection of SARS-CoV-2 infection in 4880 cases from one hospital in Wuhan, China, from Jan to Feb 2020. Clin Chim Acta 505: 172-175. https://doi.org/10.1016/j.cca.2020.03.009
[67]	Mahardika IH , Naorungroj S , Khamcharoen W , Kin S , Rodthongkum N , Chailapakul O et al (2023) Point-of-care testing (POCT) devices for DNA detection: a comprehensive review. Adv NanoBiomed Res 3: 2300058. https://doi.org/10.1002/anbr.202300058
[68]	Xu H , Xia A , Wang D , Zhang Y , Deng S , Lu W et al (2020) An ultraportable and versatile point-of-care DNA testing platform. Sci Adv 6: eaaz7445. https://doi.org/10.1126/sciadv.aaz7445
[69]	Kerr JR (2015) A review of blood diseases and cytopenias associated with human parvovirus B19 infection. Rev Med Virol 25: 224-240. https://doi.org/10.1002/rmv.1839
[70]	Yoo S-M , Jeon Y-M , Heo S-Y (2022) Electrochemiluminescence systems for the detection of biomarkers: strategical and technological advances. Biosensors (Basel) 12: 738. https://doi.org/10.3390/bios12090738
[71]	Li S , Zhang S , Shi J , Yang X , Sun H , Li Y (2022) Sensitive and non-separation 5-hydroxymethylcytosine double-stranded DNA sensing in different mice tissues: electrogenerated chemiluminescence biosensing method incorporating DNA nanomachine amplification and nanocomposite magnetic separation. Sens Actuator B-Chem 368: 132179. https://doi.org/10.1016/j.snb.2022.132179
[72]	Zhong Y , Huang L , Lin M , Zhang Z , Liu A , Lei Y (2023) A Y-shape-structured electrochemiluminescence biosensor based on carbon quantum dots and locked nucleic acid probe for microRNA determination with single-base resolution. Biosens Bioelectron 238: 115583. https://doi.org/10.1016/j.bios.2023.115583
[73]	Ai Z , Zhao M , Han D , Chen K , Xiong D , Tang H (2021) An “on-off” electrochemiluminescence immunosensor for PIVKA-II detection based on the dual quenching of CeO ₂-Au-g-C ₃N ₄ hybrids by Ag nanocubes-VB ₂ . Biosens Bioelectron 179: 113059. https://doi.org/10.1016/j.bios.2021.113059
[74]	Cai S , Li D , Luo F , Lin Z , Huang A , Qiu B (2024) An electrochemiluminescence biosensor based on target-responsive DNA hydrogel for T-2 toxin. Microchem J 205: 111231. https://doi.org/10.1016/j.microc.2024.111231
[75]	Zhang Y , Yang W , Su M , Wang B , Yuan R , Liang W (2024) A reagent-based label free electrochemiluminescence biosensor for ultrasensitive quantification of low-abundant chloramphenicol. Microchem J 198: 110124. https://doi.org/10.1016/j.microc.2024.110124
[76]	Cohen L , Walt DR (2018) Highly sensitive and multiplexed protein measurements. Chem Rev 119: 293-321. https://doi.org/10.1021/acs.chemrev.8b00257
[77]	Huang H (2018) Matrix metalloproteinase-9 (MMP-9) as a cancer biomarker and MMP-9 biosensors: recent advances. Sensors (Basel) 18: 3249. https://doi.org/10.3390/s18103249
[78]	Albright VC , Hellmich RL , Coats JR (2016) A review of cry protein detection with enzyme-linked immunosorbent assays. J Agric Food Chem 64: 2175-2189. https://doi.org/10.1021/acs.jafc.5b03766
[79]	Iha K , Inada M , Kawada N , Nakaishi K , Watabe S , Tan YH et al (2019) Ultrasensitive ELISA developed for diagnosis Diagnostics 9: 78. https://doi.org/10.3390/diagnostics9030078
[80]	Pan M , Wang Y , Wang L , Yu X , Xu L (2021) Recent advances in visual detection for cancer biomarkers and infectious pathogens. J Mater Chem B 9: 35-52. https://doi.org/10.1039/d0tb01883j
[81]	Duffy DC (2023) Digital detection of proteins. Lab Chip 23: 818-847. https://doi.org/10.1039/D2LC00783E
[82]	You M , Peng P , Xue Z , Tong H , He W , Mao P et al (2021) A fast and ultrasensitive ELISA based on rolling circle amplification. Analyst 146: 2871-2877. https://doi.org/10.1039/D1AN00355K
[83]	Wang J , Yang X , Wang X , Wang W (2022) Recent advances in CRISPR/Cas-based biosensors for protein detection. Bioengineering 9: 512. https://doi.org/10.3390/bioengineering9100512
[84]	Liu N , Liu R , Zhang J (2022) CRISPR-Cas12a-mediated label-free electrochemical aptamer-based sensor for SARS-CoV-2 antigen detection. Bioelectrochemistry 146: 108105. https://doi.org/10.1016/j.bioelechem.2022.108105
[85]	Zhao X , Wang Z , Yang B , Li Z , Tong Y , Bi Y et al (2021) Integrating PCR-free amplification and synergistic sensing for ultrasensitive and rapid CRISPR/Cas12a-based SARS-CoV-2 antigen detection. Synth Syst Biotechnol 6: 283-291. https://doi.org/10.1016/j.synbio.2021.09.007
[86]	Mu X , Li J , Xiao S , Xu J , Huang Y , Zhao S et al (2023) Peroxidase-mimicking DNA-Ag/Pt nanoclusters mediated visual biosensor for CEA detection based on rolling circle amplification and CRISPR/Cas 12a. Sens Actuators B Chem 375: 132870. https://doi.org/10.1016/j.snb.2022.132870
[87]	Qing M , Sun Z , Wang L , Du SZ , Zhou J , Tang Q et al (2021) CRISPR/Cas12a-regulated homogeneous electrochemical aptasensor for amplified detection of protein. Sens Actuators B-Chem 348: 130713. https://doi.org/10.1016/j.snb.2021.130713
[88]	Yang X , Lv J , Yang Z , Yuan R , Chai Y (2017) A sensitive electrochemical aptasensor for thrombin detection based on electroactive Co-based metal-organic frameworks with target-triggering NESA strategy. Anal Chem 89: 11636-11640. https://doi.org/10.1021/acs.analchem.7b03056
[89]	Wang W , Geng L , Zhang Y , Shen W , Bi M , Gong T et al (2024) An rolling circle amplification-assisted CRISPR/Cas12a-based biosensor for protein detection. Microchem J 200: 110370. https://doi.org/10.1016/j.microc.2024.110370
[90]	Li D , Wang D , Dong J , Wang N , Huang H , Xu H et al (2020) False-negative results of real-time reverse-transcriptase polymerase chain reaction for severe acute respiratory syndrome coronavirus 2: role of deep-learning-based CT diagnosis and insights from two cases. Korean J Radiol 21: 505-508. https://doi.org/10.3348/kjr.2020.0146
[91]	Chen S , Li M , Weng T , Wang D , Geng J (2023) Recent progress of biosensors for the detection of lung cancer markers. J Mater Chem B 11: 5715-5747. https://doi.org/10.1039/D2TB02277J
[92]	Arya SK , Bhansali S (2011) Lung cancer and its early detection using biomarker-based biosensors. Chem Rev 111: 6783-6809. https://doi.org/10.1021/cr100420s
[93]	Ahmadi A , Kabiri S , Omidfar K (2020) Advances in HbA1c biosensor development based on field effect transistors: a review. IEEE Sens J 20: 8912-8921. https://doi.org/10.1109/JSEN.2020.2987836
[94]	Crosby D , Bhatia S , Brindle KM , Coussens LM , Dive C , Emberton M et al (2022) Early detection of cancer. Science 375: eaay9040. https://doi.org/10.1126/science.aay9040
[95]	Hendrix A , Lippens L , Pinheiro C , Théry C , Martin-Jaular L , Löt-vall J et al (2023) Extracellular vesicle analysis. Nat Rev Methods Primers 3: 56. https://doi.org/10.1038/s43586-023-00240-z
[96]	Khodaparast M , Sharley D , Marshall S , Beddoe T (2024) Advances in point-of-care and molecular techniques to detect waterborne pathogens. NPJ Clean Water 7: 74. https://doi.org/10.1038/s41545-024-00368-9
[97]	Guan X , Li H , Chen L , Qi G , Jin Y (2023) Glass capillary-based nanopores for single molecule/single cell detection. ACS Sens 8: 427-442. https://doi.org/10.1021/acssensors.2c02102
[98]	Wang H , Wu T , Li M , Tao Y (2021) Recent advances in nanomaterials for colorimetric cancer detection. J Mater Chem B 9: 921-938. https://doi.org/10.1039/D0TB02163F
[99]	Shen J , Ma Z , Xu J , Xue T , Lv X , Zhu G et al (2024) Exosome isolation and detection: from microfluidic chips to nanoplasmonic biosensor. ACS Appl Mater Interfaces 16: 22776-22793. https://doi.org/10.1021/acsami.3c19396
[100]	Suea-Ngam A , Howes PD , deMello AJ (2021) An amplification-free ultra-sensitive electrochemical CRISPR/Cas biosensor for drug-resistant bacteria detection. Chem Sci 12: 12733-12743. https://doi.org/10.1039/D1SC02197D
[101]	Deusenbery C , Wang Y , Shukla A (2021) Recent innovations in bacterial infection detection and treatment. ACS Infect Dis 7: 695-720. https://doi.org/10.1021/acsinfecdis.0c00890
[102]	Bai Z , Xu X , Wang C , Wang T , Sun C , Liu S et al (2022) A comprehensive review of detection methods for Escherichia coli O157:H7 . TrAC Trends Anal Chem 152: 116646. https://doi.org/10.1016/j.trac.2022.116646
[103]	Qazi RA , Aman N , Ullah N , Jamila N , Bibi N (2024) Recent advancement for enhanced E. coli detection in electrochemical biosensors . Microchem J 196: 109673. https://doi.org/10.1016/j.microc.2023.109673
[104]	Cui R , Tang H , Huang Q , Ye T , Chen J , Huang Y et al (2024) AI-assisted smartphone-based colorimetric biosensor for visualized, rapid and sensitive detection of pathogenic bacteria. Biosens Bioelectron 259: 116369. https://doi.org/10.1016/j.bios.2024.116369
[105]	Chen Z , Ma L , Bu S , Zhang W , Chen J , Li Z et al (2021) CRISPR/Cas12a and immuno-RCA based electrochemical biosensor for detecting pathogenic bacteria. J Electroanal Chem 901: 115755. https://doi.org/10.1016/j.jelechem.2021.115755
[106]	Zhang J , Zhou M , Mao B , Huang B , Wen H , Ren J (2024) The construction of COFs functionalized CRISPR electrochemical sensor for ultrasensitive detection of bacteria by hyper-branched rolling circle amplification. Sens Actuators B Chem 409: 135610. https://doi.org/10.1016/j.snb.2024.135610
[107]	Liu Y , Li P , Cui R , Qin C , Wu L , Zhang X et al (2024) Metal-organic frameworks (MOFs) and covalent organic frameworks (COFs)-based prototyping of integrated sensing devices for robust analysis. TrAC Trends Anal Chem 174: 117678. https://doi.org/10.1016/j.trac.2024.117678
[108]	Li C , Liang Y , Feng Q (2025) An electrochemical biosensor utilizing CRISPR/Cas12a amplification for the detection of E. coli . Analyst 150: 1158-1166. https://doi.org/10.1039/D4AN01441C
[109]	Knox J , Uhlemann A-C , Lowy FD (2015) Staphylococcus aureus infections: transmission within households and the community . Trends Microbiol 23: 437-444. https://doi.org/10.1016/j.tim.2015.03.007
[110]	Lee AS , de Lencastre H , Garau J , Kluytmans J , Malhotra-Kumar S , Peschel A et al (2018) Methicillin-resistant Staphylococcus aureus . Nat Rev Dis Primers 4: 18033. https://doi.org/10.1038/nrdp.2018.33
[111]	Wang Q , Yang Q (2024) Seizing the hidden assassin: current detection strategies for Staphylococcus aureus and methicillin-resistant S. aureus . J Agric Food Chem 72: 16569-16582. https://doi.org/10.1021/acs.jafc.4c02421
[112]	Zheng L , Jin W , Xiong K , Zhen H , Li M , Hu Y (2023) Nanomaterial-based biosensors for the detection of foodborne bacteria: a review. Analyst 148: 5790-5804. https://doi.org/10.1039/D3AN01554H
[113]	Zhou B , Ye Q , Li F , Xiang X , Shang Y , Wang C et al (2022) CRISPR/Cas12a based fluorescence-enhanced lateral flow biosensor for detection of Staphylococcus aureus . Sens Actuators B Chem 351: 130906. https://doi.org/10.1016/j.snb.2021.130906
[114]	Huang L , Yuan N , Guo W , Zhang Y , Zhang W (2023) An electrochemical biosensor for the highly sensitive detection of Staphylococcus aureus based on SRCA-CRISPR/Cas12a . Talanta 252: 123821. https://doi.org/10.1016/j.talanta.2022.123821
[115]	Gao Y , Huang K , Wang F , Hou Y , Xu J , Li G (2022) Recent advances in biological detection with rolling circle amplification: design strategy, biosensing mechanism, and practical applications. Analyst 147: 3396-3414. https://doi.org/10.1039/D2AN00556E
[116]	Wei L , Wang Z , Wu L , Chen Y (2023) CRISPR/Cas12a-based magnetic relaxation switching biosensor for nucleic acid amplification-free and ultrasensitive detection of methicillin-resistant Staphylococcus aureus . Biosens Bioelectron 222: 114984. https://doi.org/10.1016/j.bios.2022.114984
[117]	Zhang Y , Yang H , Zhou Z , Huang K , Yang S , Han G (2017) Recent advances on magnetic relaxation switching assay-based nanosensors. Bioconjug Chem 28: 869-879. https://doi.org/10.1021/acs.bioconjchem.7b00059
[118]	Chen Y , Xie M (2015) A magnetic relaxation switching immunosensor for one-step detection of salbutamol based on gold nanoparticle–streptavidin conjugate. RSC Adv 5: 95401-95404. https://doi.org/10.1039/C5RA19126B
[119]	Dong Y , Wen C , She Y , Zhang Y , Chen Y , Zeng J (2021) Magnetic relaxation switching immunoassay based on hydrogen peroxide-mediated assembly of Ag@Au–Fe ₃O ₄ nanoprobe for detection of aflatoxin B1 . Small 17: 2104596. https://doi.org/10.1002/smll.202104596
[120]	Tian B , Qiu Z , Ma J , Donolato M , Hansen MF , Svedlindh P et al (2018) On-particle rolling circle amplification-based core–satellite magnetic superstructures for microRNA detection. ACS Appl Mater Interfaces 10: 2957-2964. https://doi.org/10.1021/acsami.7b16293
[121]	Zhen D , Zhang S , Yang A , Ma Q , Deng Z , Fang J et al (2024) A supersensitive electrochemical sensor based on RCA amplification-assisted “silver chain”-linked gold interdigital electrodes and CRISPR/Cas9 for the detection of Staphylococcus aureus in food . Food Chem 440: 138197. https://doi.org/10.1016/j.foodchem.2023.138197
[122]	He F , Xiong Y , Liu J , Tong F , Yan D (2016) Construction of Au-IDE/CFP10-ESAT6 aptamer/DNA-AuNPs mspqc for rapid detection of Mycobacterium tuberculosis . Biosens Bioelectron 77: 799-804. https://doi.org/10.1016/j.bios.2015.10.054
[123]	Shi X , Zhang J , He F (2019) A new aptamer/polyadenylated DNA interdigitated gold electrode piezoelectric sensor for rapid detection of Pseudomonas aeruginosa . Biosens Bioelectron 132: 224-229. https://doi.org/10.1016/j.bios.2019.02.053
[124]	Lv Z , Wang Q , Yang M (2021) Multivalent duplexed-aptamer networks regulated a CRISPR-Cas12a system for circulating tumor cell detection. Anal Chem 93: 12921-12929. https://doi.org/10.1021/acs.analchem.1c02228
[125]	Du H , Wang F , Zhang R , Yan X , Zheng J , Zhou T et al (2024) Rolling circle amplification-based self-assembly to form a “GPS-nanoconveyor” for in vitro targeted imaging and enhanced gene/chemo (CRISPR/DOX) synergistic therapy. Biomacromol 25: 4991-5007. https://doi.org/10.1021/acs.biomac.4c00415
[126]	Chieng A , Wan Z , Wang S (2024) Recent advances in real-time label-free detection of small molecules. Biosensors 14: 80. https://doi.org/10.3390/bios14020080
[127]	Deng X , Ma B , Gong Y , Li J , Zhou Y , Xu T et al (2024) Advances in aptamer-based conjugate recognition techniques for the detection of small molecules in food. Foods 13: 1749. https://doi.org/10.3390/foods13111749
[128]	Gu B , Zhang Q (2018) Recent advances on functionalized upconversion nanoparticles for detection of small molecules and ions in biosystems. Adv Sci 5: 1700609. https://doi.org/10.1002/advs.201700609
[129]	Feng W , Newbigging AM , Tao J , Cao Y , Peng H , Le C et al (2021) CRISPR technology incorporating amplification strategies: molecular assays for nucleic acids, proteins, and small molecules. Chem Sci 12: 4683-4698. https://doi.org/10.1039/d0sc06973f
[130]	Liang M , Li Z , Wang W , Liu J , Liu L , Zhu G et al (2019) A CRISPR-Cas12a-derived biosensing platform for the highly sensitive detection of diverse small molecules. Nat Commun 10: 3672. https://doi.org/10.1038/s41467-019-11648-1
[131]	Marchese S , Polo A , Ariano A , Velotto S , Costantini S , Severino L (2018) Aflatoxin B1 and M1: biological properties and their involvement in cancer development. Toxins 10: 214. https://doi.org/10.3390/toxins10060214
[132]	Shabeer S , Asad S , Jamal A , Ali A (2022) Aflatoxin contamination, its impact and management strategies: an updated review. Toxins 14: 307. https://doi.org/10.3390/toxins14050307
[133]	Abnous K , Danesh NM , Ramezani M , Alibolandi M , Nameghi MA , Zavvar TS et al (2021) A novel colorimetric aptasensor for ultrasensitive detection of aflatoxin M1 based on the combination of CRISPR-Cas12a, rolling circle amplification and catalytic activity of gold nanoparticles. Anal Chim Acta 1165: 338549. https://doi.org/10.1016/j.aca.2021.338549
[134]	Wang Z , Wei L , Ruan S , Chen Y (2023) CRISPR/Cas12a-assisted chemiluminescence sensor for aflatoxin B1 detection in cereal based on functional nucleic acid and in-pipet rolling circle amplification. J Agric Food Chem 71: 4417-4425. https://doi.org/10.1021/acs.jafc.3c00341
[135]	Zhang D , Lu S (2022) Human exposure to neonicotinoids and the associated health risks: a review. Environ Int 163: 107201. https://doi.org/10.1016/j.envint.2022.107201
[136]	Pan T , Guo W , Lu P , Hu D (2020) In situ and rapid determination of acetamiprid residue on cabbage leaf using surface-enhanced Raman scattering . J Sci Food Agric 101: 3595-3604. https://doi.org/10.1002/jsfa.10988
[137]	Yang W , Wu Y , Tao H , Zhao J , Chen H , Qiu S (2017) Ultrasensitive and selective colorimetric detection of acetamiprid pesticide based on the enhanced peroxidase-like activity of gold nanoparticles. Anal Methods 9: 5484-5493. https://doi.org/10.1039/C7AY01451A
[138]	Wang J , Wu Y , Zhou P , Yang W , Tao H , Qiu S et al (2018) A novel fluorescent aptasensor for ultrasensitive and selective detection of acetamiprid pesticide based on the inner filter effect between gold nanoparticles and carbon dots. Analyst 143: 5151-5160. https://doi.org/10.1039/C8AN01166D
[139]	Chen P , Qiao X , Liu J , Xia F , Tian D , Zhou C (2019) Dual-signaling amplification electrochemical aptasensor based on hollow polymeric nanospheres for acetamiprid detection. ACS Appl Mater Interfaces 11: 14560-14566. https://doi.org/10.1021/acsami.9b00308
[140]	Chen Y , Liang J , Xu J , Shan L , Lv J , Wu C et al (2024) Ultrasensitive paper-based photoelectrochemical biosensor for acetamiprid detection enabled by spin-state manipulation and polarity-switching. Anal Chem 96: 12262-12269. https://doi.org/10.1021/acs.analchem.4c01251
[141]	Li Y , Yang F , Yuan R , Zhong X , Zhuo Y (2022) Electrochemiluminescence covalent organic framework coupling with CRISPR/Cas12a-mediated biosensor for pesticide residue detection. Food Chem 389: 133049. https://doi.org/10.1016/j.foodchem.2022.133049
[142]	Li W , Li Y , Zhao L , Yuan R , Zhuo Y , Zhong X (2024) Triple-helix as a target converter for trace pesticide detection based on CRISPR/Cas12a-based ECL biosensor. Sens Actuators B Chem 409: 135599. https://doi.org/10.1016/j.snb.2024.135599
[143]	Zheng J , Li J , Jiang Y , Jin J , Wang K , Yang R et al (2011) Design of aptamer-based sensing platform using triple-helix molecular switch. Anal Chem 83: 6586-6592. https://doi.org/10.1021/ac201314y
[144]	Wang Y , Fang Z , Ning G , Mao S , Wu Y , Wu S et al (2019) G-quadruplex-bridged triple-helix aptamer probe strategy: a label-free chemiluminescence biosensor for ochratoxin A. Sens Actuators B Chem 298: 126867. https://doi.org/10.1016/j.snb.2019.126867
[145]	Wang Y , Zhu H , Kannan K (2019) A review of biomonitoring of phthalate exposures. Toxics 7: 21. https://doi.org/10.3390/toxics7020021
[146]	Johansson HKL , Svingen T , Fowler PA , Vinggaard AM , Boberg J (2017) Environmental influences on ovarian dysgenesis—developmental windows sensitive to chemical exposures. Nat Rev Endocrinol 13: 400-414. https://doi.org/10.1038/nrendo.2017.36
[147]	Khasin LG , Della Rosa J , Petersen N , Moeller J , Kriegsfeld LJ , Lishko PV (2020) The impact of di-2-ethylhexyl phthalate on sperm fertility. Front Cell Dev Biol 8: 426. https://doi.org/10.3389/fcell.2020.00426
[148]	Kataria A , Trasande L , Trachtman H (2015) The effects of environmental chemicals on renal function. Nat Rev Nephrol 11: 610-625. https://doi.org/10.1038/nrneph.2015.94
[149]	Braun JM (2016) Early-life exposure to EDCs: role in childhood obesity and neurodevelopment. Nat Rev Endocrinol 13: 161-173. https://doi.org/10.1038/nrendo.2016.186
[150]	Heindel JJ , Blumberg B (2019) Environmental obesogens: mechanisms and controversies. Ann Rev Pharmacol Toxicol 59: 89-106. https://doi.org/10.1146/annurev-pharmtox-010818-021304
[151]	Wang H , He B , Yan H , Liang Y , Wang J , Jin H et al (2022) Recjf exonuclease-assisted signal amplification for the sensitive detection of di(2-ethylhexyl)phthalate by a Pd@Au NB labeled electrochemical aptasensor based on Au@Ni-CoHNB/PEI-g-C ₃N ₄ . J Mater Chem C 10: 18040-18051. https://doi.org/10.1039/D2TC02602C
[152]	Dong M , Jiang D , Cao Q , Wang W , Shiigi H , Chen Z (2023) A metal-organic framework regulated graphdiyne-based electrochemiluminescence sensor with a electrocatalytic self-acceleration effect for the detection of di-(2-ethylhexyl) phthalate. Analyst 148: 4470-4478. https://doi.org/10.1039/D3AN00954H
[153]	Xiong S , Cheng J , He L , Wang M , Zhang X , Wu Z (2014) Detection of di(2-ethylhexyl)phthalate through graphene–β-cyclodextrin composites by electrochemical impedance spectroscopy. Anal Methods 6: 1736-1742. https://doi.org/10.1039/C3AY42039F
[154]	Tu D , Garza JT , Coté GL (2019) A SERS aptasensor for sensitive and selective detection of bis(2-ethylhexyl)phthalate. RSC Adv 9: 2618-2625. https://doi.org/10.1039/C8RA09230C
[155]	Liao D , Zhi J , Wang Q , Yan W , Guo Y , Han Y et al (2023) Efficient photoelectrochemical aptasensing of di-2-ethylhexyl phthalate in environmental samples based on N, S co-doped graphene quantum dots/TiO ₂ nanorods . Anal Chim Acta 1271: 341477. https://doi.org/10.1016/j.aca.2023.341477
[156]	Meng X , Huang A , Li Y , Dong X , You T (2024) Highly sensitive and selective photoelectrochemical detection of bis(2-ethylhexyl) phthalate on broad-spectrum responsive and interfacial electronic interaction induced p–n BiOI/ZnO nanoarrays heterojunction. Biosens Bioelectron 251: 116121. https://doi.org/10.1016/j.bios.2024.116121
[157]	Deng Y , Yan W , Guo Y , Wang Q , Bi Y , Dong C et al (2022) Highly sensitive and selective photoelectrochemical aptasensing of di-2-ethylhexyl phthalate based on graphene quantum dots decorated TiO ₂ nanotube arrays . J Hazard Mater 426: 128107. https://doi.org/10.1016/j.jhazmat.2021.128107
[158]	Deng Y , He R , Lu H , Guo Y , Wang Q , Xiao Y et al (2023) Visible-light driven and efficient photoelectrochemical aptasensor constructed with N-doped carbon quantum dots-decorated TiO ₂ nanorods for determination of di-2-ethylhexyl phthalate . Chem Eng J 468: 143583. https://doi.org/10.1016/j.cej.2023.143583
[159]	Yang H , Li Y , Tu C , Zhuang Y , Li Q , Li Z et al (2024) Double-enzyme active MnO ₂@BSA mediated lab-on-paper dual-modality aptasensor for di(2-ethylhexyl)phthalate . Anal Chim Acta 1287: 342135. https://doi.org/10.1016/j.aca.2023.342135
[160]	Yang H , Tu C , Zhuang Y , Li Y , Hao Y , Li Q et al (2024) Dual near-infrared AgInS ₂ and CuInS ₂ co-sensitized ZnO photoelectrode array enabled paper-based ratiometric photoelectrochemical aptasensing . Rare Met 43: 6525-6536. https://doi.org/10.1007/s12598-024-02874-8
[161]	Lu M , Zhao K , Zhang S , Cai X , Kandegama W , Chen M et al (2024) Research progress of biosensor based on organic photoelectrochemical transistor. J Agric Food Chem 72: 17746-17761. https://doi.org/10.1021/acs.jafc.4c04191
[162]	Zhang H , Zhang M , Yu Z , Zhou Y , Hu Y , Gao L et al (2024) MXene-enhanced Bi2S3/CdIn2S4 heterojunction photosensitive gate for DEHP detection in a signal-on OPECT aptamer biosensor. Anal Chem 96: 1948-1956. https://doi.org/10.1021/acs.analchem.3c04111
[163]	Zhang H , Zhang M , Zhou Y , Qiao Z , Gao L , Cao L et al (2024) Organic photoelectrochemical transistor aptasensor for dual-mode detection of DEHP with CRISPR-Cas13a assisted signal amplification. J Hazard Mater 470: 134175. https://doi.org/10.1016/j.jhazmat.2024.134175
[164]	Nuovo GJ (2008) In situ detection of precursor and mature microRNAs in paraffin embedded, formalin fixed tissues and cell preparations. Methods 44: 39-46. https://doi.org/10.1016/j.ymeth.2007.10.008
[165]	Casoli T , Stefano GD , Fattoretti P , Solazzi M , Delfino A , Biagini G et al (2003) GAP-43 mRNA detection by in situ hybridization, direct and indirect in situ RT-PCR in hippocampal and cerebellar tissue sections of adult rat brain. Micron 34: 415-422. https://doi.org/10.1016/S0968-4328(03)00038-6
[166]	McHenry A , Iyer K , Wang J , Liu C , Harigopal M (2022) Detection of SARS-CoV-2 in tissue: the comparative roles of RT-qPCR, in situ RNA hybridization, and immunohistochemistry. Expert Rev Mol Diagn 22: 559-574. https://doi.org/10.1080/14737159.2022.2085508
[167]	Lee S , Dang H , Moon J-I , Kim K , Joung Y , Park S et al (2024) SERS-based microdevices for use as in vitro diagnostic biosensors. Chem Soc Rev 53: 5394-5427. https://doi.org/10.1039/d3cs01055d
[168]	Xu K , Zhou R , Takei K , Hong M (2019) Toward flexible surface-enhanced Raman scattering (SERS) sensors for point-of-care diagnostics. Adv Sci (Weinh) 6: 1900925. https://doi.org/10.1002/advs.201900925
[169]	Asamoah BO , Uurasjärvi E , Räty J , Koistinen A , Roussey M , Peiponen K-E (2021) Towards the development of portable and in situ optical devices for detection of micro-and nanoplastics in water: a review on the current status. Polymers 13: 730. https://doi.org/10.3390/polym13050730
[170]	Xie L , Zeng H , Zhu J , Zhang Z , Sun H , Xia W et al (2022) State of the art in flexible SERS sensors toward label-free and onsite detection: from design to applications. Nano Res 15: 4374-4394. https://doi.org/10.1007/s12274-021-4017-4
[171]	Lin Z , He L (2019) Recent advance in SERS techniques for food safety and quality analysis: a brief review. Curr Opin Food Sci 28: 82-87. https://doi.org/10.1016/j.cofs.2019.10.001
[172]	Zhang D , Pu H , Huang L , Sun D-W (2021) Advances in flexible surface-enhanced Raman scattering (SERS) substrates for nondestructive food detection: fundamentals and recent applications. Trends Food Sci Technol 109: 690-701. https://doi.org/10.1016/j.tifs.2021.01.058
[173]	Han J , Wang C , Zhu L , Yang Y (2023) Emerging sensing and in situ detection technologies for the analysis of extracellular vesicle miRNAs. Adv NanoBiomed Res 4: 2300067. https://doi.org/10.1002/anbr.202300067
[174]	Feng J , Gong Y , Li Q , Yang C , An Y , Wu L (2024) In situ detection of nucleic acids in extracellular vesicles via membrane fusion. Chemistry 30: e202304111. https://doi.org/10.1002/chem.202304111
[175]	Deng R , Zhang K , Sun Y , Ren X , Li J (2017) Highly specific imaging of mRNA in single cells by target RNA-initiated rolling circle amplification. Chem Sci 8: 3668-3675. https://doi.org/10.1039/c7sc00292k
[176]	Li Z , Lau C , Lu J (2016) Effect of the concentration difference between magnesium ions and total ribonucleotide triphosphates in governing the specificity of T7 RNA polymerase-based rolling circle transcription for quantitative detection. Anal Chem 88: 6078-6083. https://doi.org/10.1021/acs.analchem.6b01460
[177]	Chen F , Xue J , Zhang J , Bai M , Yu X , Fan C et al (2020) Differentiated visualization of single-cell 5-hydroxymethylpyrimidines with microfluidic hydrogel encoding. J Am Chem Soc 142: 2889-2896. https://doi.org/10.1021/jacs.9b11393
[178]	Tian M , Zhang R , Li J (2023) Emergence of CRISPR/Cas9-mediated bioimaging: a new dawn of in-situ detection. Biosens Bioelectron 232: 115302. https://doi.org/10.1016/j.bios.2023.115302
[179]	Liu Y , Li S , Zhang L , Zhao Q , Li N , Wu Y (2020) A sensitive and specific method for microRNA detection and in situ imaging based on a CRISPR-Cas9 modified catalytic hairpin assembly . RSC Adv 10: 28037-28040. https://doi.org/10.1039/d0ra03603j
[180]	Zhou X , Tang W , Zhang Y , Deng A , Guo Y , Qian L (2024) Liposome–exosome hybrids for in situ detection of exosomal miR-1246 in breast cancer. Analyst 149: 403-409. https://doi.org/10.1039/d3an01600e
[181]	Zhang T , Xie Z , Zheng X , Liang Y , Lu Y , Zhong H et al (2024) CRISPR-Cas12a powered hybrid nanoparticle for extracellular vesicle aggregation and in-situ microRNA detection. Biosens Bioelectron 245: 115856. https://doi.org/10.1016/j.bios.2023.115856
[182]	Kim H , Gu C , Mustfa SA , Martella DA , Wang C , Wang Y et al (2023) CRISPR/Cas-assisted nanoneedle sensor for adenosine triphosphate detection in living cells. ACS Appl Mater Interfaces 15: 49964-49973. https://doi.org/10.1021/acsami.3c07918
[183]	Zhou Y , Che S , Wang Z , Zhang X , Yuan X (2024) Primer exchange reaction assisted CRISPR/Cas9 cleavage for detection of dual microRNAs with electrochemistry method. Microchim Acta 191: 502. https://doi.org/10.1007/s00604-024-06548-1
[184]	Meng L , Wang L , Sun Z , Mu G , Li Z , Wu J (2025) Selective in situ analysis of hepatogenic exosomal microRNAs via virus-mimicking multifunctional magnetic vesicles. Adv Healthc Mater 14: e2404981. https://doi.org/10.1002/adhm.202404981
[185]	Li J , Sun D (2022) Molecularly imprinted ratiometric fluorescence nanosensors. Langmuir 38: 13305-13312. https://doi.org/10.1021/acs.langmuir.2c01925
[186]	Park JY , Kricka LJ (2007) Prospects for nano- and microtechnologies in clinical point-of-care testing. Lab Chip 7: 547-549. https://doi.org/10.1039/b702667f
[187]	Herrmann L , Breuer J , Duc TN , Thomé N , Ghazaani F , Kamhieh-Milz S et al (2024) Comparison of the diagnostic accuracy of the Pluslife mini dock RHAM technology with Abbott ID now and Cepheid GenXpert: a retrospective evaluation study. Sci Rep 14: 13978. https://doi.org/10.1038/s41598-024-64406-9
[188]	Zahavi M , Rohana H , Azrad M , Shinberg B , Peretz A (2022) Rapid SARS-CoV-2 detection using the Lucira™ check it COVID-19 test kit. Diagnostics 12: 1877. https://doi.org/10.3390/diagnostics12081877
[189]	Lévesque S , Beauchemin S , Vallée M , Longtin J , Jacob-Wagner M , Dumaresq J et al (2022) Evaluation of water gargle samples for SARS-CoV-2 detection using Abbott ID NOW COVID-19 assay. J Med Virol 94: 4522-4527. https://doi.org/10.1002/jmv.27847
[190]	Tsang HF , Leung WMS , Chan LWC , Cho WSC , Wong SCC (2021) Performance comparison of the Cobas® Liat® and Cepheid® GeneXpert® systems on SARS-CoV-2 detection in nasopharyngeal swab and posterior oropharyngeal saliva. Expert Rev Mol Diagn 21: 515-518. https://doi.org/10.1080/14737159.2021.1919513
[191]	Kellner MJ , Koob JG , Gootenberg JS , Abudayyeh OO , Zhang F (2019) SHERLOCK: nucleic acid detection with CRISPR nucleases. Nat Protoc 14: 2986-3012. https://doi.org/10.1038/s41596-019-0210-2
[192]	Chen JS , Ma E , Harrington LB , Da Costa M , Tian X , Palefsky JM et al (2018) CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science 360: 436-439. https://doi.org/10.1126/science.aar6245
[193]	Sena-Torralba A , Álvarez-Diduk R , Parolo C , Piper A , Merkoçi A (2022) Toward next generation lateral flow assays: integration of nanomaterials. Chem Rev 122: 14881-14910. https://doi.org/10.1021/acs.chemrev.1c01012
[194]	Ivanov AV , Safenkova IV , Zherdev AV , Dzantiev BB (2022) DIRECT 2: a novel platform for a CRISPR-Cas12-based assay comprising universal DNA-IgG probe and a direct lateral flow test. Biosens Bioelectron 208: 114227. https://doi.org/10.1016/j.bios.2022.114227
[195]	Ding R , Long J , Yuan M , Zheng X , Shen Y , Jin Y et al (2021) CRISPR/Cas12-based ultra-sensitive and specific point-of-care detection of HBV. Int J Mol Sci 22: 4842. https://doi.org/10.3390/ijms22094842
[196]	Zhao X , Wang Z , Zhang H , Liu J , Wu W , Yu L et al (2024) Highly sensitive one-pot isothermal assay combining rolling circle amplification and CRISPR/Cas12a for aflatoxin B1 detection. Anal Chem 96: 18070-18078. https://doi.org/10.1021/acs.analchem.4c03798
[197]	Liu M , Li X , Xu J , Zhou S , Deng L , Men D et al (2024) Sensitive detection of CaMV35S based on exponential rolling circle amplification reaction and CRISPR/Cas12a using a portable 3D-printed visualizer. Microchem J 205: 111313. https://doi.org/10.1016/j.microc.2024.111313
[198]	Gong Y , Zheng Y , Jin B , You M , Wang J , Li X et al (2019) A portable and universal upconversion nanoparticle-based lateral flow assay platform for point-of-care testing. Talanta 201: 126-133. https://doi.org/10.1016/j.talanta.2019.03.105
[199]	Everitt ML , Tillery A , David MG , Singh N , Borison A , White IM (2021) A critical review of point-of-care diagnostic technologies to combat viral pandemics. Anal Chim Acta 1146: 184-199. https://doi.org/10.1016/j.aca.2020.10.009
[200]	Takeuchi Y , Akashi Y , Kato D , Kuwahara M , Muramatsu S , Ueda A et al (2021) Diagnostic performance and characteristics of anterior nasal collection for the SARS-CoV-2 antigen test: a prospective study. Sci Rep 11: 10519. https://doi.org/10.1038/s41598-021-90026-8
[201]	Jeong Y , Kook Y-M , Lee K , Koh W-G (2018) Metal enhanced fluorescence (MEF) for biosensors: general approaches and a review of recent developments. Biosens Bioelectron 111: 102-116. https://doi.org/10.1016/j.bios.2018.04.007
[202]	Xu L , Li D , Ramadan S , Li Y , Klein N (2020) Facile biosensors for rapid detection of COVID-19. Biosens Bioelectron 170: 112673. https://doi.org/10.1016/j.bios.2020.112673
[203]	Xu L , Shoaie N , Jahanpeyma F , Zhao J , Azimzadeh M , Al Jamal KT (2020) Optical, electrochemical and electrical (nano)biosensors for detection of exosomes: a comprehensive overview. Biosens Bioelectron 161: 112222. https://doi.org/10.1016/j.bios.2020.112222
[204]	Qin X , Liu J , Zhang Z , Li J , Yuan L , Zhang Z et al (2021) Microfluidic paper-based chips in rapid detection: current status, challenges, and perspectives. Trends Anal Chem 143: 116371. https://doi.org/10.1016/j.trac.2021.116371
[205]	Hai X , Li Y , Zhu C , Song W , Cao J , Bi S (2020) DNA-based label-free electrochemical biosensors: from principles to applications. Trends Analyt Chem 133: 116098. https://doi.org/10.1016/j.trac.2020.116098
[206]	Abbas N , Song S , Chang M-S , Chun M-S (2023) Point-of-care diagnostic devices for detection of Escherichia coli O157:H7 using microfluidic systems: a focused review . Biosensors (Basel) 13: 741. https://doi.org/10.3390/bios13070741
[207]	Oliveira LS , Goel S , Amreen K , Lucena-Silva N , Oliveira MDL , Andrade CAS (2024) Microfluid biosensor for detection of HPV in patient scraping samples: determining E6 and E7 oncogenes. Bioelectrochemistry 160: 108795. https://doi.org/10.1016/j.bioelechem.2024.108795
[208]	Chen H , Cai S , Luo J , Liu X , Ou L , Zhang Q et al (2024) Colorimetric biosensing assays based on gold nanoparticles functionalized/combined with non-antibody recognition elements. Trends Analyt Chem 173: 117654. https://doi.org/10.1016/j.trac.2024.117654
[209]	Hao X , Liu Y , Wen Y , Chen W , Ramadan S , Jin N et al (2025) Two-pot ready-to-use reagents achieved quadruple-signal amplification for the ultra-sensitive biosensing of Salmonella typhimurium in foods . Biosens Bioelectron 286: 117627. https://doi.org/10.1016/j.bios.2025.117627
[210]	Ye X , Wu H , Liu J , Xiang J , Feng Y , Liu Q (2024) One-pot diagnostic methods based on CRISPR/Cas and Argonaute nucleases: strategies and perspectives. Trends Biotechnol 42: 1410-1426. https://doi.org/10.1016/j.tibtech.2024.06.009
[211]	Lesinski JM , Moragues T , Mathur P , Shen Y , Paganini C , Bezzinge L et al (2024) In situ complexation of sgRNA and Cas12a improves the performance of a one-pot RPA-CRISPR-Cas12 assay. Anal Chem 96: 10443-10450. https://doi.org/10.1021/acs.analchem.4c01777
[212]	Janghorban M , Aradanas I , Kazemi S , Ngaju P , Pandey R (2022) Recent advances, opportunities, and challenges in developing nucleic acid integrated wearable biosensors for expanding the capabilities of wearable technologies in health monitoring. Biosensors (Basel) 12: 986. https://doi.org/10.3390/bios12110986
[213]	Liu R , He Y , Lan T , Zhang J (2021) Installing CRISPR-Cas12a sensors in a portable glucose meter for point-of-care detection of analytes. Analyst 146: 3114-3120. https://doi.org/10.1039/d1an00008j
[214]	Nguyen PQ , Soenksen LR , Donghia NM , Angenent-Mari NM , de Puig H , Huang A et al (2021) Wearable materials with embedded synthetic biology sensors for biomolecule detection. Nat Biotechnol 39: 1366-1374. https://doi.org/10.1038/s41587-021-00950-3
[215]	Politza AJ , Nouri R , Guan W (2023) Digital CRISPR systems for the next generation of nucleic acid quantification. Trends Analyt Chem 159: 116917. https://doi.org/10.1016/j.trac.2023.116917
[216]	Anwar N , Jiang G , Wen Y , Ahmed M , Zhong H , Ao S et al (2024) Evaluating the potential of two-dimensional materials for innovations in multifunctional electrochromic biochemical sensors: a review. Moore More 1: 12. https://doi.org/10.1007/s44275-024-00013-0
[217]	Ren Q , Jia S , Li J , He L , Xu Y , Huang H et al (2025) Integrated plasmonic ruler using terahertz multi-BIC metasurface for digital biosensing. Moore More 2: 5. https://doi.org/10.1007/s44275-025-00027-2

Recent advances in CRISPR- and RCA-based biosensing chips and devices for POCT and in situ detection

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