[1]马贤参,常新昊,司文喆.CRISPR-Cas12a系统在病原体检测领域的最新研究进展[J].现代检验医学杂志,2026,41(02):192-197.[doi:10.3969/j.issn.1671-7414.2026.02.032]
　MA Xiancan,CHANG Xinhao,SI Wenzhe.Recent Advances in the CRISPR-Cas12a System for Pathogen Detection[J].Journal of Modern Laboratory Medicine,2026,41(02):192-197.[doi:10.3969/j.issn.1671-7414.2026.02.032]
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CRISPR-Cas12a系统在病原体检测领域的最新研究进展()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
192-197
栏目:
综述
出版日期:
2026-03-15

文章信息/Info

Title:
Recent Advances in the CRISPR-Cas12a System for Pathogen Detection
文章编号:
1671-7414（2026）02-192-06
作者:
马贤参a，常新昊a，司文喆a,b,c
北京大学第三医院a.检验科；b.心血管分子生物学与调节肽重点实验室；c.血管稳态与重构全国重点实验室，北京 100191
Author(s):
MA Xiancana, CHANG Xinhaoa, SI Wenzhea,b,c
a. Department of Clinical Laboratory; b. Key Laboratory of Cardiovas-cular Molecular Biology and Regulatory Peptides; c. State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University Third Hospital, Beijing 100191, China
关键词:
成簇规律的间隔短回文重复序列-关联蛋白12a系统; 病原体; 核酸检测
分类号:
R-331；R446
DOI:
10.3969/j.issn.1671-7414.2026.02.032
文献标志码:
A
摘要:
成簇规律间隔短回文重复序列-关联蛋白12a(CRISPR-Cas12a)体系是一种源自细菌的自然免疫防御机制,因其高特异度、高灵敏度、无需依赖昂贵设备、操作简便以及成本效益显著等优点,在核酸分析领域引起广泛的关注。CRIS-PR-Cas12a系统能够与扩增技术、生物传感器、拉曼光谱、微流控等相关技术联用,从而提高检测系统的灵敏度,减少交叉污染,缩短检测时间,极大拓宽了其应用场景。近年来该技术被广泛应用于病毒、细菌、支原体等病原体核酸检测中。该文主要介绍了CRISPR-Cas12a系统在上述病原体检测中的研究进展,深入探讨了其优势和局限性,旨在推动该技术在病原体的快速识别、精准诊断及现场筛查场景中的深度应用与迭代升级的实践思考。
Abstract:
The clustered regulatory interspaced short palindromic repeat - associated protein 12a (CRISPR-Cas12a) system is a natural immune defense mechanism derived from bacteria, which has garnered significant attention in the field of nucleic acid analysis due to its high specificity, high sensitivity, equipment-free operation, simplicity and remarkable cost-effectiveness. The CRISPR-Cas12a system can be integrated with related technologies such as amplification techniques, biosensors, Raman spec-troscopy, and microfluidics to enhance the detection sensitivity of the system, reduce cross contamination, shorten the testing time, and broaden the application scenarios. In recent years, the CRISPR-Cas12a technology has been widely applied in nucleic acid detection for pathogens such as viruses, bacteria and mycoplasmas. This article primarily reviews the research advances of the CRISPR-Cas12a system in the aforementioned pathogen detection applications, provides an in-depth analysis of its advantag-es and limitations, and proposes practical strategies to deepen its implementation and iterative refinement in rapid pathogen iden-tification, precise diagnosis, and on-site screening scenarios.

参考文献/References:

[1] KONG X Q, WANG Y J, FANG Z X, et al. False-positive results of SARS-CoV-2 RT-PCR in oropharyngeal swabs from vaccinators [J]. Frontiers in Medicine, 2022, 9:847407.
[2] RAHBARI R, MORADI N, ABDI M. RRT-PCR for SARS-CoV-2: analytical considerations [J]. Clinica Chimica Acta, 2021, 516: 1-7.
[3] 赵亚楠,曹啟新,闫彦,等.CRISPR/Cas13系统用于病原体检测的研究进展[J].检验医学,2023,38(8):800-804. ZHAO Y N, CAO Q X, YAN Y, et al. Research prog-ress of CRISPR/Cas13 system for pathogen detec-tion[J]. Laboratory Medicine, 2023, 38(8): 800-804.
[4] 张咪咪.CRISPR-Cas技术在病原体核酸检测中的研究进展[J].国际检验医学杂志,2023,44(10):1259-1265. ZHANG M M. Research progress of CRISPR-Cas systems in nucleic acid detection of pathogens[J]. International Jour-nal of Laboratory Medicine, 2023, 44(10): 1259-1265.
[5] MAKAROVA K S, WOLF Y I, ALKHNBASHI O S, et al. An updated evolutionary classification of CRISPR-Cas systems[J]. Nature Reviews Microbiology, 2015, 13(11): 722-736.
[6] 党生,张帅,翟景波.CRISPR/Cas12a系统:核酸检测的多功能工具[J].生物化学与生物物理进展, 2024, 51(4): 785-796. DANG S, ZHANG S, ZHAI J B. The versatile tool: CRIS-PR/Cas12a system for nucleic acid detection[J]. Progress in Biochemistry and Biophysics, 2024, 51(4): 785-796.
[7] PAUL B, MONTOYA G. CRISPR-Cas12a: functional overview and applications[J]. Biomedical Journal, 2020, 43(1): 8-17.
[8] 周桓,邵艳娜,王涓,等.基于CRISPR/Cas技术的核酸检测研究进展[J].微生物学报,2021,61(12):3856-3869. ZHOU H, SHAO Y N, WANG J, et al. Research progress of nucleic acid detection based on CRISPR/Cas technology[J]. Acta Microbiologica Sinica, 2021, 61(12): 3856-3869.
[9] 冯万有,蒙丽娜,陈春,等.CRISPR/Cas系统分类和病原体检测的研究进展[J].生物技术, 2022, 32(2):258-267, 251. FENG W Y, MENG L N, CHEN C, et al. Classification of CRISPR/Cas system and application in pathogen de-tection[J]. Biotechnology, 2022, 32(2): 258-267, 251.
[10] WANG J Y, PAUSCH P, DOUDNA J A. Structural biology of CRISPR-Cas immunity and genome editing enzymes[J]. Nature Reviews Microbiology, 2022, 20(11): 641-656.
[11] WANG Y H, YANG T M, LIU G F, et al. Application of CRISPR/Cas12a in the rapid detection of pathogens [J]. Clinica Chimica Acta, 2023, 548: 117520.
[12] 王雅轩,朱晓雁,苏建荣.CRISPR/Cas系统在病原体检测方面的研究进展[J].中国人兽共患病学报, 2021, 37(9): 839-844. WANG Y X, ZHU X Y, SU J R. Research progress on the CRISPR/Cas system in pathogen detection[J]. Chi-nese Journal of Zoonoses, 2021, 37(9): 839-844.
[13] YAO R L, LIU D, JIA X, et al. CRISPR-Cas9/Cas12a biotechnology and application in bacteria [J]. Synthetic and Systems Biotechnology, 2018, 3(3): 135-149.
[14] 周建,任雪梅,王馨,等.CRISPR/Cas核酸检测系统的研究进展和未来挑战[J].检验医学, 2024, 39(6):608-614. ZHOU J, REN X M, WANG X, et al. Research progress and challenges on CRISPR/Cas nucleic acid diagnosis system[J]. Laboratory Medicine, 2024, 39(6): 608-614.
[15] CHEN J S, MA E B, HARRINGTON L B, et al. CRISPR-Cas12a target binding unleashes indiscrimi-nate single-stranded DNase activity[J]. Science, 2018, 360(6387): 436-439.
[16] LI S Y, CHENG Q X, WANG J M, et al. CRISPR-Cas12a-as-sisted nucleic acid detection [J]. Cell Discovery, 2018, 4: 20.
[17] 陈逸凡.CRISPR/Cas技术在病原体检测中的应用[J].检验医学与临床,2022,19(7):981-985. CHEN Y F. Application of CRISPR/Cas technology in pathogen detection[J]. Laboratory Medicine and Clinic, 2022, 19(7): 981-985.
[18] DAI Y F, SOMOZA R A, WANG L, et al. Exploring the trans-cleavage activity of CRISPR-Cas12a (cpf1) for the development of a universal electrochemical biosensor[J]. Angewandte Chemie (International ed. in English), 2019, 58(48): 17399-17405.
[19] 陈思,林昱坤,宋春燕,等.基于CRISPR/Cas系统的核酸生物传感器[J].生物化学与生物物理进展, 2023, 50(8): 1782-1796. CHEN S, LIN Y K, SONG C Y, et al. CRISPR/Cas-based biosensors for nucleic acid detection[J]. Progress in Bio-chemistry and Biophysics, 2023, 50(8): 1782-1796.
[20] LIU P F, ZHAO K R, LIU Z J, et al. Cas12a-based electrochemiluminescence biosensor for target amplifi-cation-free DNA detection [J]. Biosensors and Bioelec-tronics, 2021, 176: 112954.
[21] NOURI R Z, JIANG Y Q, LIAN X L, et al. Se-quence-specific recognition of HIV-1 DNA with sol-id-state CRISPR-Cas12a-assisted nanopores (SCAN) [J]. ACS Sensors, 2020, 5(5): 1273-1280.
[22] WANG H M, SU A L, BAO C X, et al. A CRISPR/Ca-s12a-SERS platform for amplification-free detection of Afri-can swine fever virus genes [J]. Talanta, 2024, 267: 125225.
[23] LIANG J J, TENG P J, XIAO W, et al. Application of the amplification-free SERS-based CRISPR/Cas12a platform in the identification of SARS-CoV-2 from clinical sam-ples[J]. Journal of Nanobiotechnology, 2021, 19(1): 273.
[24] WANG M X, CHANG J T, HAN Y X, et al. A CRISPR/Cas12a-based platform for rapid on-site bovine viral diarrhea virus diagnostics[J]. Journal of Integrative Ag-riculture, 2024, 23(8): 2872-2876.
[25] 董铮,赵振兴,范奇璇,等.基于RPA-CRISPR/Cas12a的番茄花叶病毒可视化检测方法的建立[J].植物保护, 2024, 50(4): 235-241. DONG Z, ZHAO Z X, FAN Q X, et al. Establishment of RPA-CRISPR/Cas12a-based visual detection of Tomato mosaic virus[J]. Plant Protection, 2024, 50(4): 235-241.
[26] 于晶雪,韦珊珊,覃绍敏,等.基于CRISPR/Cas12a-RT-RAA的猪繁殖与呼吸综合征病毒快速检测方法的建立[J].中国畜牧兽医, 2024, 51(8): 3237-3246. YU J X, WEI S S, QIN S M, et al. Establishment of a rapid detection method for porcine reproductive and respiratory syndrome virus based on CRISPR/Cas12a-RT-RAA[J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(8): 3237-3246.
[27] QIAN W D, WANG X F, WANG T, et al. Development of RPA-Cas12a-fluorescence assay for rapid and reli-able detection of human bocavirus 1[J]. Animal Models and Experimental Medicine, 2024, 7(2): 179-188.
[28] WANG X, RAO Q, LU Z R, et al. Rapid and sensitive Cas13a/Cas12a-based one-pot dual-target strategy to detect monkeypox virus and its co-infected viruses[J]. Science Bulletin, 2023, 68(24): 3142-3148.
[29] 吴永彬,李凌.CRISPR/Cas系统在新型冠状病毒肺炎快速诊断中的应用[J].现代检验医学杂志, 2022, 37(3): 1-5. WU Y B, LI L. Application of CRISPR/Cas systems in the rapid diagnosis of coronavirus disease 2019[J]. Journal of Modern Laboratory Medicine, 2022, 37(3): 1-5.
[30] GAYET R V, DE PUIG H, ENGLISH M A, et al. Creat-ing CRISPR-responsive smart materials for diagnostics and programmable cargo release[J]. Nature Protocols, 2020, 15(9): 3030-3063.
[31] ZHANG L, WANG H L, YANG S, et al. High-through-put and integrated CRISPR/Cas12a-Based molecular diagnosis using a deep learning enabled microfluidic system[J]. ACS Nano, 2024, 18(35): 24236-24251.
[32] LIU Y J, LIU H, YU G L, et al. One-tube RPA-CRISPR Cas12a/Cas13a rapid detection of methicillin-resistant Staphylococcus aureus [J]. Analytica Chimica Acta, 2023, 1278: 341757.
[33] XU J H, BAI X R, ZHANG X L L, et al. Development and application of DETECTR-based rapid detection for pathogenic Bacillusanthracis [J]. Analytica Chimica Acta, 2023, 1247: 340891.
[34] ZHANG W J, QU H, WU X, et al. Rapid, sensitive, and user-friendly detection of Pseudomonas aeruginosa using the RPA/CRISPR/Cas12a system[J]. BMC Infec-tious Diseases, 2024, 24(1): 458.
[35] ZHU L S, LIANG Z D, XU Y T, et al. Ultrasensitive and rapid visual detection of Escherichia coli O157:H7 based on RAA-CRISPR/Cas12a system[J]. Biosensors, 2023, 13(6): 659.
[36] JIA N, WANG C H, LIU X M, et al. A CRISPR-Ca-s12a-based platform for ultrasensitive rapid highly spe-cific detection of Mycobacterium tuberculosis in clin-ical application [J]. Frontiers in Cellular and Infection Microbiology, 2023, 13: 1192134.
[37] LIU J H, CHEN J H, WU D, et al. CRISPR-/Cas12a-me-diated liposome-amplified strategy for the surface-en-hanced raman scattering and naked-eye detection of nucle-ic acid and application to food authenticity screening[J]. Analytical Chemistry, 2021, 93(29): 10167-10174.
[38] ZHUANG J W, ZHAO Z Y, LIAN K, et al. SERS-based CRISPR/Cas assay on microfluidic paper analytical devices for supersensitive detection of pathogenic bacteria in foods [J]. Biosensors & Bioelectronics, 2022, 207: 114167.
[39] JIA N, ZHOU J, XIAO F, et al. A CRISPR-Ca-s12a-based platform for ultrasensitive, rapid, and highly specific detection of Mycoplasma pneumonia in clinical application [J]. Frontiers in Bioengineering and Bio-technology, 2023, 11: 1022066.
[40] CHEN J L, HUANG Y E, XIAO B, et al. Development of a RPA-CRISPR-Cas12a assay for rapid, simple, and sensitive detection of Mycoplasma hominis [J]. Fron-tiers in Microbiology, 2022, 13: 842415.
[41] LI S, WANG X C, YU Y H, et al. Establishment and application of a CRISPR-Cas12a-based RPA-LFS and fluorescence for the detection of Trichomonas vaginal-is[J]. Parasites&Vectors, 2022, 15(1): 350.
[42] HUANG F Q, LI X, ZHOU Y L, et al. Optimization of CRISPR/Cas12a detection assay and its application in the detection of Echinococcus granulosus [J]. Veteri-nary Parasitology, 2024, 331: 110276.
[43] MAO Z F, WANG X J, CHEN R P, et al. Upconver-sion-mediated CRISPR-Cas12a biosensing for sensitive detection of ochratoxin A [J]. Talanta, 2022, 242: 123232.
[44] WU Z H, SUN D W, PU H B. CRISPR/Cas12a and G-qua-druplex DNAzyme-driven multimodal biosensor for visual detection of Aflatoxin B1 [J]. Spectrochimica Acta Part A: Mo-lecular and Biomolecular Spectroscopy, 2023, 302: 123121.
[45] WANG B, WANG R, WANG D Q, et al. Cas12aVDet:A CRISPR/Cas12a-based platform for rapid and visual nucleic acid detection[J]. Analytical Chemistry, 2019, 91(19): 12156-12161.

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备注/Memo

备注/Memo:
基金项目： 北京市科技新星计划（20220484090）；北京市科技新星交叉合作课题（20230484442）；北京市自然科学基金（7232206）；血管稳态与重构全国重点实验室研究基金资助(2024-VHR-SY-13)。
作者简介： 马贤参（2001-），男，在读博士生，研究方向：病原体检测和分子诊断新技术，E-mail：2110117118@bjmu.edu.cn。
常新昊（2003-），男，在读本科生，专业：医学检验技术，E-mail：2210117140@stu.pku.edu.cn，并列第一作者。
通讯作者：司文喆（1987-），女，副研究员，硕士生导师，研究方向：分子诊断新技术研究，E-mail：wenzhesi@bjmu.edu.cn。
更新日期/Last Update: 2026-03-15