微生物学报

Recognition element	Advantages	Disadvantages	References
Antibodies	High specificity, high selectivity, mature technology	Prone to false positives, large molecular weight, poor stability, high cost	[21]
Aptamers	High specificity, good stability, low immunogenicity, scalable in vitro production	Complex screening process, lack of universal systematic evolution of ligand by exponential enrichment (SELEX) standards, requires stability improvement	[22]
Bacteriophages	Strong specificity, low cost, simple operation, protease-insensitive	Immature biosensor integration, requires technical development	[23]
Molecularly imprinted polymers (MIP)	High stability, simple preparation, low cost	Weak binding affinity, low selectivity, challenging large-scale synthesis, susceptible to matrix interference	[24]
Lectins	Broad-spectrum binding, unique glycosyl recognition capability, high stability	Limited selectivity, insufficient affinity and specificity	[25]

Recognition element	Advantages	Disadvantages	References
Antibodies	High specificity, high selectivity, mature technology	Prone to false positives, large molecular weight, poor stability, high cost	[21]
Aptamers	High specificity, good stability, low immunogenicity, scalable in vitro production	Complex screening process, lack of universal systematic evolution of ligand by exponential enrichment (SELEX) standards, requires stability improvement	[22]
Bacteriophages	Strong specificity, low cost, simple operation, protease-insensitive	Immature biosensor integration, requires technical development	[23]
Molecularly imprinted polymers (MIP)	High stability, simple preparation, low cost	Weak binding affinity, low selectivity, challenging large-scale synthesis, susceptible to matrix interference	[24]
Lectins	Broad-spectrum binding, unique glycosyl recognition capability, high stability	Limited selectivity, insufficient affinity and specificity	[25]

Sensor type	Electrode	Functional material system	Recognition element	Detection sample	Limit of detection (CFU/mL)	Linear range (CFU/mL)	References
Amperometric	Screen-printed three-electrode system (SPE)	Gold nanoparticles (GNPs)	Aptamer	Water	60	6.0×10¹-6.0×10⁷	[28]
Gold electrode	AuNPs/Super P/Cu-ZrMOF	Antibody, aptamer	Urine	2	10¹-10⁷	[29]
Carbon screen-printed electrode (CSPE)	MIL-101(Cr)/MWCNT/AgNPs/c-g-C₃N₄	Aptamer	Serum	1	10¹-10⁷	[30]
GCE	AuNPs	Aptamer-MIP	Serum	1	10¹-10⁷	[31]
GCE	CCLP/AuNPs	Monoclonal antibody	Water	9×10²	10¹-10⁷	[32]
GCE	HZIFs-8/Fc-GO	Aptamer	Urine	1	1.2×10¹-1.2×10⁷	[33]
Au-SPEs	MB	Aptamer	Water	8	0-10¹⁰	[34]
GCE	CoFe₂O₄/AgNPs	Pyocyanin	Apple, beef, water, chicken, fish, egg, soil, milk	4.0×10^-3	1-23	[35]
Impedimetric	GCE	AgNPs	Aptamer	Serum	33	10²-10⁷	[36]
GCE	NCs/Glu	Aptamer	Serum	3	10¹-10⁷	[37]
GCE	CNCNF	Aptamer	Serum	1	10¹-10⁷	[38]
ECL	GCE	Carboxyl graphene	Phage	Milk, glucose injection, urine	56	1.4×10²-1.4×10⁶	[39]
Piezoelectric	Gold (Au) interdigital electrode (Au IDE)	Polyadenylated DNA/MB	Aptamer	Buﬀer, blood	9 (buﬀer), 52 (blood)	8.1×10¹-8.1×10⁵ (buﬀer), 1.9×10²-1.0×10⁶ (blood)	[40]

Sensor type	Electrode	Functional material system	Recognition element	Detection sample	Limit of detection (CFU/mL)	Linear range (CFU/mL)	References
Amperometric	Screen-printed three-electrode system (SPE)	Gold nanoparticles (GNPs)	Aptamer	Water	60	6.0×10¹-6.0×10⁷	[28]
Gold electrode	AuNPs/Super P/Cu-ZrMOF	Antibody, aptamer	Urine	2	10¹-10⁷	[29]
Carbon screen-printed electrode (CSPE)	MIL-101(Cr)/MWCNT/AgNPs/c-g-C₃N₄	Aptamer	Serum	1	10¹-10⁷	[30]
GCE	AuNPs	Aptamer-MIP	Serum	1	10¹-10⁷	[31]
GCE	CCLP/AuNPs	Monoclonal antibody	Water	9×10²	10¹-10⁷	[32]
GCE	HZIFs-8/Fc-GO	Aptamer	Urine	1	1.2×10¹-1.2×10⁷	[33]
Au-SPEs	MB	Aptamer	Water	8	0-10¹⁰	[34]
GCE	CoFe₂O₄/AgNPs	Pyocyanin	Apple, beef, water, chicken, fish, egg, soil, milk	4.0×10^-3	1-23	[35]
Impedimetric	GCE	AgNPs	Aptamer	Serum	33	10²-10⁷	[36]
GCE	NCs/Glu	Aptamer	Serum	3	10¹-10⁷	[37]
GCE	CNCNF	Aptamer	Serum	1	10¹-10⁷	[38]
ECL	GCE	Carboxyl graphene	Phage	Milk, glucose injection, urine	56	1.4×10²-1.4×10⁶	[39]
Piezoelectric	Gold (Au) interdigital electrode (Au IDE)	Polyadenylated DNA/MB	Aptamer	Buﬀer, blood	9 (buﬀer), 52 (blood)	8.1×10¹-8.1×10⁵ (buﬀer), 1.9×10²-1.0×10⁶ (blood)	[40]

Sensor type	Signal system (amplifier/ reporter/quencher)	Recognition element	Detection sample	Limit of detection (CFU/mL)	Linear range (CFU/mL)	References
SERS	AuNPs (amplifier), 4-MBA (reporter)	Aptamer	Water, chicken meat	20	10²-10⁷	[49]
Fe₃O₄@Au (amplifier), DTNB (reporter)	Cecropin 1, antibody	Urine	12 cell/mL	/	[50]
AuNSs (amplifier), DTNB (reporter)	WGA, antibody	Urine	5	10¹-10⁶	[51]
AuNPs (amplifier), DTNB (reporter)	Single-guide RNA, CRISPR/dCas9	Urine	1	1-10⁶	[52]
Au (amplifier)	/	Blood	5×10³	/	[53]
Fluorescent	TPE (reporter)	Aptamer	Water, orange juice, milk	1×10²	10²-10⁸	[54]
CDs (reporter), GO (quencher)	Aptamer	Water	9	10¹-10⁷	[55]
PDA-PEI copolymer dots (reporter)	Dual aptamer	Milk, orange juice, popsicle	1	10¹-10⁷	[56]
HCR (amplifier), FAM (reporter), BHQ-1 (quencher)	Aptamer	Urine	37	10²-10⁷	[57]
FAM (reporter)	DNAzyme (PAE-1)	Water, pekoe tea, peach juice, hawthorn juice	1.2	/	[58]
FAM-cDNA (reporter), GOQDs (quencher)	Aptamer	Water, orange juice, popsicle	1×10²	1.28×10³-2.0×10⁷	[59]
SPR	MG-NPA (amplifier)	Aptamer	/	30 CFU/assay	/	[60]
Au nanotriangle array (amplifier)	Aptamer	/	10	10¹-10³	[61]

Sensor type	Signal system (amplifier/ reporter/quencher)	Recognition element	Detection sample	Limit of detection (CFU/mL)	Linear range (CFU/mL)	References
SERS	AuNPs (amplifier), 4-MBA (reporter)	Aptamer	Water, chicken meat	20	10²-10⁷	[49]
Fe₃O₄@Au (amplifier), DTNB (reporter)	Cecropin 1, antibody	Urine	12 cell/mL	/	[50]
AuNSs (amplifier), DTNB (reporter)	WGA, antibody	Urine	5	10¹-10⁶	[51]
AuNPs (amplifier), DTNB (reporter)	Single-guide RNA, CRISPR/dCas9	Urine	1	1-10⁶	[52]
Au (amplifier)	/	Blood	5×10³	/	[53]
Fluorescent	TPE (reporter)	Aptamer	Water, orange juice, milk	1×10²	10²-10⁸	[54]
CDs (reporter), GO (quencher)	Aptamer	Water	9	10¹-10⁷	[55]
PDA-PEI copolymer dots (reporter)	Dual aptamer	Milk, orange juice, popsicle	1	10¹-10⁷	[56]
HCR (amplifier), FAM (reporter), BHQ-1 (quencher)	Aptamer	Urine	37	10²-10⁷	[57]
FAM (reporter)	DNAzyme (PAE-1)	Water, pekoe tea, peach juice, hawthorn juice	1.2	/	[58]
FAM-cDNA (reporter), GOQDs (quencher)	Aptamer	Water, orange juice, popsicle	1×10²	1.28×10³-2.0×10⁷	[59]
SPR	MG-NPA (amplifier)	Aptamer	/	30 CFU/assay	/	[60]
Au nanotriangle array (amplifier)	Aptamer	/	10	10¹-10³	[61]

多类生物传感技术在铜绿假单胞菌检测中的研究进展

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黄洁洁 , 郑华 ^*

微生物学报 | 综述 2025,65(11): 4800-4816

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微生物学报 | 综述 2025, 65(11): 4800-4816

多类生物传感技术在铜绿假单胞菌检测中的研究进展

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黄洁洁, 郑华^*

作者信息

广西医科大学生命科学研究院，广西南宁

Research progress in the detection of Pseudomonas aeruginosa by multi-class biosensing technologies

Jiejie HUANG, Hua ZHENG^*

Affiliations

Life Sciences Institute, Guangxi Medical University, Nanning, Guangxi, China

出版时间: 2025-11-04 doi: 10.13343/j.cnki.wsxb.20250327

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摘要

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铜绿假单胞菌(Pseudomonas aeruginosa, PA)是引发医院感染的重要病原菌。它能在各种极端环境中生存，且具有高度抗生素耐药性，可攻击免疫功能低下人群，进而引发严重感染，是导致临床患者死亡的重要原因。因此，实现PA的快速诊断对于控制感染至关重要。传统检测技术，如平板法、免疫分析法和核酸法等在准确性和敏感性方面表现优异，但存在成本较高、检测时间长的问题。近年来，新型生物传感检测技术通过集成多样化的生物识别元件和信号增强策略实现了对PA的超敏、精准检测，为PA的快速识别、治疗及控制提供了高效、便捷且经济的解决方案。本文系统综述了基于电化学、光学、CRISPR/Cas12a系统和磁性纳米颗粒磁分离的生物传感技术的原理、应用进展及面临的挑战，并对未来研究方向进行了展望，以促进PA快速检测技术的创新与临床应用。

关键词

铜绿假单胞菌 / 生物传感技术 / 电化学 / 光学 / CRISPR/Cas12a / 磁性纳米颗粒

Abstract

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Pseudomonas aeruginosa (PA) is a major cause of hospital-acquired infections. It can survive in a variety of extreme environments, and is highly resistant to antibiotics. PA can attack immunocompromised populations to cause severe infections, being a major cause of mortality in clinical practice. Therefore, rapid diagnosis of PA is critical for infection control. Conventional detection techniques such as plate assays, immunoassays, and nucleic acid assays have excellent accuracy and sensitivity, while they are costly and time-consuming. In recent years, novel biosensing technologies have achieved ultrasensitive and precise detection of PA by integrating various biorecognition elements and signal enhancement strategies, providing an efficient, convenient, and cost-effective solution for the rapid identification, treatment, and control of PA. In this paper, we systematically review the principles, application progress, and challenges of magnetic separation biosensing technologies based on electrochemical, optical, CRISPR/Cas12a system, and magnetic nanoparticles, and provide future research directions, aiming to promote the innovation and clinical application of the technologies for rapid detection of PA.

Key words

Pseudomonas aeruginosa / biosensing technology / electrochemistry / optics / CRISPR/Cas12a / magnetic nanoparticles

引用本文

黄洁洁, 郑华. 多类生物传感技术在铜绿假单胞菌检测中的研究进展. 微生物学报, 2025 , 65 (11) : 4800 -4816 . DOI: 10.13343/j.cnki.wsxb.20250327

Jiejie HUANG, Hua ZHENG. Research progress in the detection of Pseudomonas aeruginosa by multi-class biosensing technologies[J]. Acta Microbiologica Sinica, 2025 , 65 (11) : 4800 -4816 . DOI: 10.13343/j.cnki.wsxb.20250327

正文

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铜绿假单胞菌(Pseudomonas aeruginosa, PA)是一种革兰氏阴性的杆状细菌，隶属于假单胞菌属。它广泛分布于自然界的水体、土壤和空气中，还能在各种极端环境下生存，并通过形成生物膜增强自身抵抗能力^[1]。作为医院感染的重要病原体之一，PA可引发尿路感染^[2]、呼吸道感染^[3]、血液感染、皮肤感染^[4]等多种感染，是导致囊性纤维化(cystic fibrosis, CF)患者肺部感染、呼吸功能下降甚至死亡的重要原因^[5]，同时也是器官移植和大面积烧伤患者患败血症的主要原因^[6]。PA的致病机理复杂，涉及多种毒力因子，如外膜蛋白、脂多糖、外毒素A、鞭毛、IV型菌毛、吡嗪蓝素和生物膜等^[7]。这些毒力因子协同作用抑制宿主免疫应答，增强其在宿主体内的存活和传播能力。值得注意的是，在这些毒力因子中生物膜是PA抵御抗生素和宿主防御的关键，其形成受PA的群体感应系统调控，本质上是由细菌及其自身产生的胞外聚合物(extracellular polymeric substance, EPS)基质组成的复杂聚集体，多糖、细胞外DNA等基质成分占其组成的90%，为黏附和包裹菌体提供了保护作用^[8-9]。这一保护屏障的存在使得抗菌剂无法完全消灭细菌，导致反复感染的临床后果^[10]，甚至能在10-100 CFU/mL的极低浓度下引发感染^[11]。此外，PA还能通过药物外排泵、改变药物作用靶点、产生抗菌药物灭活酶等机制提高耐药能力^[12]，堪称名副其实的“超级细菌”。在世界卫生组织(World Health Organization, WHO) 2017年发布的抗生素耐药菌新抗生素研发优先清单中，PA被归类为“关键”级别^[13]，这意味着针对PA开发新型抗生素极为紧迫。鉴于PA感染的多样性及严重的耐药性问题，快速、准确的病原学诊断对于指导临床治疗、控制感染传播具有重要意义。

目前已开发出多种检测PA的方法，主要有平板培养法、滤膜法、聚合酶链反应法、酶联免疫法、质谱法等^[14-16]。这些方法提供了从传统培养到分子诊断的多样化手段，具有高特异性和高灵敏度的优势，但也存在一定的局限性。传统平板培养法作为金标准，检测周期较长；滤膜法的滤膜性能评价标准不统一，检验效率差异较大；分子诊断方法可能存在交叉反应，且价格昂贵、需要专业人员操作，其经济性和实用性均不满足快速检测要求。因此，急需新型检测手段对PA进行实时监控和预防。随着分子生物学和纳米技术的发展，微小型传感器检测技术因其经济、便携、功耗低、响应速度快等优点受到广泛关注，并逐渐被开发应用于PA检测^[17]。本文详细综述各类新型PA传感检测技术的基本原理、应用及面临的挑战，以期为克服传统检测方法的局限性、推动PA检测技术的创新发展提供科学依据。

1 生物传感技术

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生物传感技术是一种利用生物分子识别元件对特定分析物进行识别，并将目标分析物的存在和浓度转换为可测量信号的技术。其雏形可追溯至1967年，Updieke等^[18]成功研制出第一个酶生物传感器，为生物传感技术的蓬勃发展奠定了基础，开启了新的发展篇章。识别元件(如抗体、适配体、噬菌体、分子印迹聚合物、凝集素等)是生物传感技术的核心组成部分，能特异性识别并捕获目标物，将其存在及浓度转变为可测量的物理、化学信号，从而对目标分析物进行定性和定量分析^[19-20]，识别元件相关信息见表1。生物传感技术因其多功能性、集成性、快速响应和经济便捷等特点广泛应用于医疗诊断和食品安全领域^[26]。目前，已研发出多种新型的生物传感技术应用于病原菌的检测，其突破性检测手段革新了传统检测模式，已成为公共卫生安全领域的热点研究方向。

1.1 电化学生物传感技术

电化学生物传感技术是一种将生物识别元件与电化学信号转换器相结合的分析技术。其原理是利用生物识别元件捕获目标分析物后在电极界面上产生电化学信号，再通过相应的信号转换器将该信号转变为可识别的电信号并进行分析处理，通过监测这些电信号的变化来定量检测目标分析物^[27]。该技术是生物传感技术领域中最典型、最具代表性的一类，具有高灵敏度和高特异性的优点。PA菌体本身能被生物识别元件特异性捕获，从而导致电化学信号变化。此外，PA在生长过程中通过代谢活动产生或消耗特定的化学物质，如绿脓菌素等，电化学传感器同样能对其代谢物质进行检测并产生电化学信号，该信号与目标物浓度呈线性关系，由此可实现对PA的快速、灵敏、定量检测^[28]。表2总结了不同电化学生物传感器在PA检测中的应用。

1.1.1 电流型

电流型生物传感器是一种通过测量电流变化来定量分析目标物浓度的技术。该技术以电流信号作为输出，其强度与目标物浓度成正比，从而实现高灵敏检测^[41]。优质的修饰材料是有效增强电化学生物传感器性能的关键。碳基纳米材料与贵金属因具有良好的机械性能、高电化学活性和大比表面积的优势常被用于修饰工作电极^[42]。Huang等^[43]以还原氧化石墨烯(reduced graphene oxide, rGO)、多壁碳纳米管(multi-walled carbon nanotube, MWCNT)两类碳基材料，协同贵金属金纳米颗粒(gold nanoparticles, AuNPs)构建复合物体系制备电化学传感器，实现了茶饮料中铅含量的高效测定；Zheng等^[44]在黄曲霉毒素B1检测中同样利用AuNPs作为电子传导增强剂提升了电化学传感器的灵敏度。同样的策略也适用于PA传感器，Zhang等^[29]通过合成具有高催化活性的铜-锆金属有机框架(Cu-Zr metal organic framework, Cu-ZrMOF)，结合适配体和DNA构建信号放大探针，采用AuNPs和炭黑材料(Super P)修饰电极构建传感平台，利用AP抗体和适配体捕获细菌，进而通过Cu-ZrMOF催化H₂O₂的分解改变电流信号实现对PA的定量检测，检出限为2 CFU/mL。Abedi等^[30]设计了一种MIL-101(Cr)/MWCNT修饰碳丝网印刷电极(carbon screen-printed electrode, CSPE)的电流型适配体传感器，该传感器的特点在于采用了三明治型结构，类似于酶联免疫吸附测定(enzyme-linked immunosorbent assay, ELISA)试剂盒^[45]。MIL-101(Cr)/MWCNT修饰电极增加了有效表面积，适配体F23捕获探针固定在电极表面，引入银纳米颗粒(silver nanoparticles, AgNPs)和二维石墨相氮化碳复合物(c-g-C₃N₄)作为信号标签与F23适配体识别，形成夹层式结构，该方法能实现双信号放大银氧化电流信号，实现对PA的超灵敏检测，对人血清样本检出限低至1 CFU/mL^[30]。分子印迹聚合物(molecular imprinted polymer, MIP)凭借其可定制的识别空腔正成为电化学传感器领域的研究热点。Ma等^[46]开发的新型便携光电化学传感器以MIP为识别单元，实现了对唾液中尿酸的超灵敏检测。MIP的抗干扰能力与实际样本检测的可行性验证同样适用于病原微生物检测，Sarabaegi等^[31]建立了一种结合MIP和适配体传感的方法来测量PA，通过电聚合技术促使电极表面形成MIP，并通过孵育PA结合影响DPV峰电流变化来实现定量检测PA。MIP具有快速识别、高稳定性、低成本的独特优势，但是MIP的结合亲和力较弱是导致其不能广泛应用在实际现场检测的原因之一^[47]。Krithiga等^[32]利用钙离子交联果胶(calcium cross-linked pectin, CCLP)和AuNPs复合材料(CCLP/AuNPs)修饰电极，并基于PA的单克隆抗体-抗原反应产生的电化学信号变化来定量检测PA，该传感器整个检测过程仅需15 min，检测限为9×10² CFU/mL。Shahrokhian等^[33]设计了一种基于中空多孔沸石咪唑酸盐框架-8 (hollow porous zeolitic imidazolate framework-8, HZIF-8)和氧化石墨烯负载茂烯(ferrocene-graphene oxide, Fc-GO)的电化学传感器，用于快速、灵敏地检测PA，检出限为1 CFU/mL。Maghsoomi等^[34]开发了一种基于亚甲蓝(methylene blue, MB)的新型电化学适配体传感器，该传感器利用细菌与适配体结合的构象变化来改变电极表面电荷分布，从而量化PA浓度，检出限为8 CFU/mL。Shobana等^[35]采用钴铁氧体掺杂银纳米颗粒(CoFe₂O₄/AgNPs)复合材料修饰电极，提高电极对绿脓菌素的电催化活性，通过监测其电化学行为来指示细菌的存在和浓度，该传感器对PA具有4.0×10^-3 CFU/mL的超低检出限。

1.1.2 阻抗型

阻抗型传感器的工作原理是通过监测电极与电解质界面的阻抗变化来反映目标分析物的存在和浓度。其具有无标记检测的优势，无需使用荧光或放射性标记物，减少了标记过程的复杂性和潜在的生物活性干扰，还具备快速响应、高灵敏度和高选择性的特点，已广泛应用于PA检测^[48]。AgNPs的电化学性能与AuNPs同样出色，Roushani等^[36]通过电沉积AgNPs修饰玻碳电极(glassy carbon electrode, GCE)，将氨基化的适配体固定化并共价连接到AgNP/GCE表面，建立了一种新型的阻抗电化学生物传感器，PA与适配体结合阻碍了电极表面与[Fe(CN)₆]^(3-/4-)电化学探针之间的电子转移，导致电荷转移阻抗增加，从而可定量分析PA浓度；该传感器是第一个用于PA检测的阻抗传感器，具有良好的特异性、重复性，对血清样品的检出限为33 CFU/mL。Sarabaegi等^[37]使用纳米壳聚糖颗粒(nano-sized chitosan particles, NCs)修饰电极构建了一个新型的电化学适配体传感器，利用戊二醛(glutaraldehyde, Glu)共价固定适配体在NCs/GCE电极表面，结合PA时引起电极表面阻抗的变化，通过阻抗信号响应反映PA浓度；该传感器对PA的检测限低至3 CFU/mL，具有良好的选择性和重复性。Sarabaegi等^[38]还制备了一种新型中空碳纳米胶囊基氮掺杂碳纳米纤维(hollow carbon nanocapsules-based nitrogen-doped carbon nanofibers, CNCNF)修饰电极，其特殊结构能为捕获细菌提供高的比表面积并提升传感器的信号放大和传导，极大提升传感器对目标分子的捕获效率和灵敏度；PA与连接在CNCNF上的适配体结合时导致电极表面电荷转移电阻增加，从而实现PA检测；该阻抗传感器展现出对PA的快速、灵敏检测能力，检出限低至1 CFU/mL，在10¹-10⁷ CFU/mL线性范围内对PA具有良好阻抗响应。

1.1.3 其他类型

除了常见的阻抗型和电流型电化学传感器外，压电传感器和电化学发光(electrochemiluminescence, ECL)传感器也因其独特优势和应用潜力被开发应用于PA的快速检测。Yue等^[39]以噬菌体PaP1为识别元件，研制了一种新型无标记ECL生物传感器，该传感器的原理是基于噬菌体尾部纤维和基板与细菌细胞壁的相互作用实现的，当PA存在时会形成非导电的噬菌体/细菌复合物，鲁米诺作为ECL试剂活性分子扩散受阻，中断了界面电子转移，ECL信号随PA浓度线性下降，从而可以推断样品中PA的存在和浓度；整个检测过程可在30 min内完成，检测灵敏度为56 CFU/mL。该传感器对PA的检测是在无标签模式下进行的，是一种友好便捷的检测方案。Shi等^[40]开发了一种基于磁珠/适配体/聚腺苷酸化DNA偶联的“三明治”压电传感器，聚腺苷酸化DNA与适配体部分互补，当PA存在时竞争性与适配体结合，聚腺苷酸化DNA被取代释放并吸附到金叉指电极上，导致多通道串联式压电石英晶体传感器频率的敏感信号移位变化，以此实现对PA的定量检测；与传统的电化学传感器相比，该方法无须标记和复杂的电极修饰，能直接通过频率变化实现高灵敏度和高特异性检测，并且成功应用于血液样品中PA的选择性检测，检出限为52 CFU/mL。这些新型传感器不仅扩展了电化学传感器的应用范围，而且在灵敏度、选择性和操作便捷性等方面也展现出了各自的优势，为未来开发更高效的PA生物传感器提供新的参考。

1.2 光学生物传感技术

光学生物传感技术是一种通过捕捉光信号的变化来感知被测物质的存在或变化的技术，其原理是借助生物识别单元与目标物质的特异性相互作用，引发光学性质的改变，如荧光增减、光谱特征峰和吸收峰的位移等，这些变化可被光学元件捕捉并转换为相应信号，通过检测这些光学信号的变化来实现对目标物质的定性或定量分析^[25]。光学生物传感技术具有高灵敏度、快速响应和低成本检测等优点，已被广泛应用于病原菌检测。表3总结了不同光学生物传感技术在PA检测中的应用。

1.2.1 表面增强拉曼散射型

表面增强拉曼散射(surface-enhanced Raman scattering, SERS)是一种利用金属纳米颗粒表面的增强效应来提高拉曼散射信号的技术。因其具备高灵敏度以及能够提供分子结构信息的能力在细菌检测领域备受关注。其基本原理是通过在金属纳米颗粒表面形成局域表面等离子体共振，从而实现拉曼散射信号的显著增强；这种增强效应可达到分子级别的分辨率，使得SERS技术能够检测到极低浓度的分子，甚至单个分子^[62]。金属纳米颗粒，如金、银等，因其优异的光学性质和高拉曼散射截面常被用作SERS基底材料。Wu等^[49]通过制备2种尺寸的AuNPs分别固定PA的互补DNA (complementary DNA, cDNA)和适配体，建立了具有SERS信号和颜色信号的探针用于识别PA；该双模式传感器利用SERS信号的减弱和比色信号的增强2种变化实现了对PA的高灵敏度和高选择性检测，在SERS模式下检测限为20 CFU/mL，用时仅需2 h。Chen等^[50]开发了一种基于抗菌肽功能化磁性标签的超高灵敏度SERS生物传感器，联合侧流层析分析(lateral flow assay, LFA)同时检测PA和大肠杆菌(Escherichia coli)O157:H7，该传感器利用链亲和素与生物素系统将抗菌肽(cecropin 1, CP1)修饰在Fe₃O₄@Au表面，获得具有广谱捕获特性的Fe₃O₄@Au-CP1纳米颗粒，该纳米颗粒能对尿液样本中的目标细菌外膜进行有效特异性结合，并利用Fe₃O₄的磁性实现快速富集；当细菌在LFA试纸条上与特定抗体结合，可快速产生视觉结果，同时测试线处产生的SERS信号被用于定量分析PA浓度；该传感器对尿液中的PA和E. coli O157:H7的检出限分别为12 cell/mL和16 cell/mL。Cheng等^[51]构建了一种磁辅助双识别的SERS生物传感器，用于同时检测PA和金黄色葡萄球菌，该传感器以抗体修饰的磁纳米颗粒作为捕获探针，小麦凝集素(wheat-germ agglutinin, WGA)修饰金纳米星(gold nanostars, AuNSs)作为SERS信号标签，形成类似于三明治的“磁纳米颗粒/细菌/SERS标签”夹心结构，最后通过检测三明治结构携带拉曼分子巯基苯甲酸和5,5′-二硫代双(2-硝基苯甲酸) [5,5′-dithiobis-(2-nitrobenzoic acid), DTNB]的特征信号强度定量获得细菌浓度；该研究是首个提出WGA和SERS标签组合的研究，实现了对PA的高灵敏度检测，检测限低至5 CFU/mL。Jiang等^[52]利用不同的拉曼分子修饰AuNPs，结合相应的巯基化DNA探针分别对金黄色葡萄球菌、PA和E. coli O157:H7的DNA进行特异性识别；经成簇规律间隔短回文重复序列(clustered regularly interspaced short palindromic repeats, CRISPR)/相关蛋白(deactivated CRISPR associated protein, dCas9)系统识别目的基因，并引导功能化AuNPs在其自组装形成SERS热点，利用智能手机集成的手持拉曼光谱仪检测，整个过程可在50 min内完成，对PA的检测限低至1 CFU/mL。Cheng等^[53]开发了一种结合SERS技术的微流控检测平台用于快速检测血液中的病原体，PA作为检测目标之一，通过混合电动力机制能够在大约3 min内被有效浓缩在SERS活性粗糙电极的停滞区域，同时排除血细胞，检出限为5×10³ CFU/mL。该平台的优势在于检测时间极短，而且能在复杂血液样本中有效分离细菌和血细胞并进行准确检测。

1.2.2 荧光型

荧光光谱法是光谱法中的一个重要分支。荧光检测是一种利用荧光物质在特定波长的光激发下发出荧光信号的技术，其基本原理是通过荧光标记物与目标物特异性结合，荧光物质受到光量子激发后会迅速发射出具有不同特性的另一量子，荧光信号被接收进而对目标物实现定量分析^[63]。Yin等^[54]将四苯基乙烯(tetraphenylethylene, TPE)衍生物与适配体结合形成3种不同的荧光探针，分别检测金黄色葡萄球菌、大肠杆菌和PA。该传感器以Fe₃O₄@SiO₂作为基体材料制备MIPs，在磁场下对细菌进行磁分离纯化和鉴定，结合智能手机与外置透镜形成荧光传感器，现场实时定量测量荧光细菌的数量，传感器灵敏度为1×10² CFU/mL，检测时间仅需40 min。在各类发光纳米材料中，发光碳点(carbon dots, CDs)因其优异的光学性能和易于大规模合成的优点走进大众视野。Wang等^[55]开发了一种基于适配体/CDs/氧化石墨烯(graphene oxide, GO)的荧光检测系统用于检测PA，适配体结合CDs形成荧光探针，通过π-π堆叠附着于GO表面，由于荧光共振能量转移(fluorescence resonance energy transfer, FRET)效应，GO猝灭了碳点的荧光，导致背景荧光信号降低；相反，当PA存在时与适配体特异性结合，不再与GO接触，FRET效应被解除，荧光信号恢复并加强，从而定量检测PA，检测限为9 CFU/mL。相较于单适配体，双适配体特异性结合更具优势。Zhong等^[56]制备了一种聚多巴胺-聚乙烯亚胺(polydopamine-polyethyleneimine, PDA-PEI)共聚物点，结合双适配体形成荧光探针捕获PA，经特异性捕获后分离细菌和探针的结合物，用380 nm和530 nm的激发/发射波长测量离心后上清液荧光强度变化，可以定量PA的存在和浓度。该策略由于构建了双适配体，对PA具有更高的亲和力，检测限低至1 CFU/mL，整个检测过程可在1.5 h内完成。Xie等^[57]提出了一种基于双链DNA分支迁移诱导的杂交链式反应(hybridization chain reaction, HCR)和DNA酶反馈回路的荧光生物传感器，该传感器利用PA与其适配体的特异性结合触发双链DNA分支迁移，产生2个DNA“Y”型结构B1A1和B2A2；B1A1激活DNA酶反馈回路，通过切割C1和C2产生更多“Y”型结构，实现循环放大；而B2A2触发HCR，打开含荧光报告基团6-羧基荧光素(6-carboxyfluorescein, FAM)和淬灭基团黑洞淬灭剂-1 (black hole quencher-1, BHQ-1)的DNA发夹结构，导致荧光信号减弱从而实现定量检测PA；该生物传感器具有检测临床尿液样本的潜力，在条件优化下检测限能达到37 CFU/mL。Qin等^[58]筛选了一种名为PAE-1的DNA酶，能对PA的细胞外产物CEM-PA表现出高切割率和强特异性；通过荧光素FAM标记底物设计了一种荧光生物传感器，当PA存在时分泌CEM-PA并激活PAE-1，随后PAE-1切割荧光标记底物，释放荧光信号；该传感器的检测限低至1.2 CFU/mL，且能在10 min内完成检测。Gao等^[59]基于石墨烯氧化物量子点(graphene oxide quantum dots, GOQDs)构建了一种新型荧光传感器，其原理是利用荧光素标记cDNA与适配体杂交，使荧光GOQDs猝灭，当PA存在时与适配体特异结合，FAM-cDNA从GOQDs表面解吸从而恢复荧光，实现PA高灵敏检测，检测限为1×10² CFU/mL。

1.2.3 表面等离子体共振型

表面等离子体共振(surface plasmon resonance, SPR)是光与金属表面自由电子相互作用产生的光学现象。当光波以特定角度入射到金属表面时，光的能量可以激发金属表面的自由电子，形成一种称为“表面等离子体”的集体振荡态，当这种振荡态的频率与入射光的频率匹配时会发生共振，导致光的折射率显著降低，通过监测生物分子在金属表面吸附引起的这种折射率变化可实现对目标分子的无标记检测^[64]。Yoo等^[60]构建了一种适配体功能化的局部SPR传感器，能同时检测包括PA在内的3种不同细菌；该传感器通过多点金帽纳米颗粒阵列(multispot gold-capped nanoparticle array, MG-NPA)芯片构建，当目标细菌与适配体结合时引起局域表面等离子体共振峰(localized surface plasmon resonance peak, LSPR)的强度变化，从而实现对细菌的无标记检测。Hu等^[61]开发了一个能检测全细胞PA菌株PAO1的局部SPR传感平台，传感器表面使用了生物素修饰的聚乙二醇、聚乙二醇1:3比例自组装，结合中性亲和素进行修饰，加入适配体后能形成一种夹层式的超稳生物识别层；PAO1菌株通过与特异性适配体结合被拉至金纳米三角形阵列表面，引起LSPR峰的偏移，以此检测目标菌株；该传感器可实现PA的单细胞水平检测，检测限为10 CFU/mL，检测时间约为3 h，具有灵敏度高、选择性好的优点。

1.3 基于CRISPR/Cas12a系统的生物传感技术

CRISPR是一种在细菌和古菌基因组中存在的特殊结构DNA区域，作为适应性防御机制在免疫系统中起着重要作用^[65-67]。联合其相关蛋白(CRISPR/associated protein, Cas)构成的CRISPR/Cas系统作为一种强大的基因编辑工具已广泛用于核酸检测领域。其中，Cas12a (也称Cpf1)作为该系统的重要成员，是一种由RNA引导的核酸内切酶。该蛋白大约含有1 200-1 500个氨基酸，隶属于2类V-A CRISPR系统的一部分，能够对目标DNA序列进行特异性识别并进行切割^[68]。基于CRISPR/Cas12a系统的生物传感器主要由信号放大、信号识别以及信号输出3个部分组成，其快速、直观的信号读取是提高检测的关键。基于CRISPR/Cas12a系统的信号读取主要是通过识别目标物后Cas12a旁切裂解荧光标记的单链DNA报告分子来释放荧光信号，除此之外，为了提升检测性能还开发出了比色的直观信号读取方式，在此详述荧光信号、比色信号输出检测。

1.3.1 荧光信号输出

基于荧光信号的读取是最为常见的一种检测方式，具有直观、简便的特点，且操作无需复杂设备，能在短时间内快速对目标物进行检测。Yang等^[69]基于重组酶聚合酶扩增(recombinase polymerase amplification, RPA)技术和CRISPR/Cas12a系统构建了一个用于PA灵敏检测的平台，该平台以PA的lasB基因作为目标基因设计特异性CRISPR RNA (crRNA)，利用RPA技术对lasB基因进行扩增，Cas12a蛋白在crRNA的引导下发挥核酸酶活性对目标基因切割，同时被激活后进一步切割带有6-FAM荧光标记的单链DNA报告分子，此时FAM荧光标记与淬灭基团分离，释放出荧光信号，利用Probit回归分析定量计算出该平台最低检出限为15.9 CFU /mL。Liu等^[70]同样构建了一个结合RPA技术和CRISPR/Cas12a系统的PA快检传感平台，RPA技术对目标基因进行等温扩增，结合CRISPR/Cas12a系统对PA进行检测可在30 min内完成，通过实时荧光分析仪监测信号来判断样品中是否存在PA，该平台非常适合于临床快速诊断和现场检测。Xiao等^[71]结合变性泡介导的链置换扩增(denaturation bubble-mediated strand exchange amplification, SEA)技术、CRISPR/Cas12a系统和噬菌体扩增开发了一种新型的活菌检测法；使用PA靶向噬菌体感染目标菌株以产生大量噬菌体实现初步扩增，再利用SEA进行二次扩增，当CRISPR/Cas12a复合体识别到目标DNA序列时Cas12a被激活并切割单链DNA导致荧光素进入激发状态，荧光信号被捕获；利用该检测平台能在3.5 h内正确区分所有人工加标的阳性样本和阴性样本，验证了其在复杂实际样品中的稳定性和准确性。腙类化合物作为一类重要的有机合成中间体，近年来也被应用于与其他技术结合，将信号放大，进一步提升病原菌检测的灵敏度和特异性。Sheng等^[72]基于腙化学介导CRISPR/Cas12a系统针对PA实现特异性定量检测，其原理设计是利用PA的16S rRNA基因中特定序列的短DNA片段作为探针，结合肼基修饰的激活链片段TS1-NHNH₂形成杂交体，特异性识别PA后释放TS1，并利用互补碱基配对诱导的邻近效应，使TS1与醛基修饰的激活链片段TS2-CHO通过腙键连接起来，形成完整的TS1/TS2激活链，随后Cas12a/crRNA被激活以特异性地切割荧光标记的单链DNA-FAM，以此产生荧光信号；该方法最低检测限为24 CFU/mL。Xu等^[73]基于CRISPR/Cas12a系统构建了ECL和荧光的双模生物传感器。铱(III)配合物作为双模探针，磁性发光单元Fe₃O₄@SiO₂@Ir/SiO₂实现信号调控；目标PA激活Cas12a反式切割活性，特异性裂解单链DNA-猝灭基团，释放铱配合物的发光信号，ECL与荧光模式检测限分别达73 fmol/L和0.126 pmol/L。该策略通过CRISPR系统与双模信号协同放大，实现了DNA超灵敏检测。Wang等^[74]开发了可变色的CRISPR/Cas12a系统比率测定法，该策略利用RPA技术扩增基因并利用Cas12a的反式切割活性切断生物发光共振能量转移报告基因，破坏荧光素酶与受体的能量转移，导致生物发光颜色由绿变蓝；结合比率测量和鲁棒传感器可以使用智能手机可视化检测金黄色葡萄球菌和PA的原子摩尔基因组DNA。

1.3.2 比色信号输出

比色信号分析是基于吸光度或RGB值变化来定量分析目标物质的方法，其基本原理是目标物质与特定试剂发生反应生成有色化合物，颜色的深浅与目标物质的浓度成正比，通过比较或测量颜色强度来推断目标物的存在和浓度。Mukama等^[75]报道了一种适用于PA现场快检的侧流生物传感器(lateral flow biosensor, LFB)检测平台，通过测流试纸条进行比色信号检测直接肉眼读出结果；该平台结合环介导等温扩增(loop-mediated isothermal amplification, LAMP)技术和CRISPR/Cas12系统，利用LAMP技术对PA的酰基转移酶特异性基因进行等温扩增，快速生成大量目标DNA；随后，扩增产物与Cas12蛋白和crRNA结合，激活Cas12的旁观者效应，切割生物素标记的单链DNA报告基因，切割后的报告基因无法在LFB的测试线上产生比色信号，从而实现对PA的可视化检测。整个检测过程无需复杂的DNA提取和纯化步骤，具有单拷贝的灵敏度和高特异性。Hu等^[76]通过整合自引物辅助链延伸和CRISPR/Cas12a系统比色显色实现对PA的灵敏检测，该方法以适配体F23为检测探头，设计了一种能特异性结合F23的自引物，利用单链DNA激活Cas12a的横向切割活性，随后Cas12a切割与银离子(Ag⁺)螯合的适配体序列，释放的Ag⁺氧化底物产生颜色变化，根据吸光度与目标菌株浓度之间的相关性实现PA定量检测；该方法对比传统依靠DNA杂交检测显色策略相比具有更高的效率，检测限为21 CFU/mL；同时，该策略的实用性也在临床样本中也得到了有效验证，通过比较该方法与传统菌落计数法对人工加标PA的血清样本定量检测，两者结果高度一致，表明该策略建立的方法有望能成为PA检测的替代方法。

1.4 基于磁性纳米颗粒的磁分离生物传感技术

磁性纳米颗粒(magnetic nanoparticles, MNPs)是一种尺寸在1-100 nm之间的磁性纳米材料^[77]，通常由铁、钴、镍、锌、铜、锰等磁性元素及其他化合物组成，具有在外部磁场作用下响应并定向移动聚集的超顺磁性^[78]。因其独特的物理化学性质和优异的生物相容性，它已成为现代生物医药和食品安全领域的重要工具^[79]。近年来，MNPs凭借其卓越的磁分离和富集能力在病原菌检测领域受到广泛关注。通过对MNPs表面进行功能化修饰引入特异性抗体或核酸适配体等，可借助其高亲和力实现对目标病原菌的高效捕获与分离。根据识别元件的不同识别机制可分为免疫识别元件和核酸识别元件。

1.4.1 免疫识别元件

免疫识别元件主要利用抗体、抗原、肽段及相关蛋白作为识别单元，通过目标分子与其特异性结合来实现高特异性检测。这种识别机制具有高选择性和多功能性，适用于检测复杂样本中的特定目标物。Liu等^[80]开发了一种基于功能化MNPs亲和捕获技术，结合基质辅助激光解吸电离质谱分析的快检平台用于鉴定PA，该研究利用免疫组件鸽卵清蛋白(pigeon ovalbumin, POA)作为亲和探针，并修饰Fe₃O₄@Al₂O₃NPs，通过POA表面的GGalα(1→4)Gal单元与PA外膜上的半乳糖亲和性凝集素特异性结合，实现对目标细菌的高效捕获和分离。Alhogail等^[81]设计了一种基于MNPs的生物传感器，以NH2-Ahx-Gly-Gly-Gly-Ahx-Cys肽段作为底物，N端通过Ahx连接子与MNPs共价结合，C端通过Cys与金表面形成自组装单层，将底物固定在金传感器表面，当PA存在并分泌LasA蛋白酶时会切割底物中的Gly-Gly键，届时底物/MNPs复合物从金表面释放并被外部磁铁吸引，暴露出金表面的金色，从而实现PA检测。实验通过对20株临床分离PA菌株测试，结果均为阳性，该传感器在1 min内的检出限为1×10² CFU/mL，可适用于PA蛋白酶的定性和半定量检测。磁性量子点(magnetic quantum dots, Mag@QDs)也被归类为MNPs的一种特殊类型，这种材料通常结合了量子点和MNPs的特性。Tu等^[82]制备了一种WGA修饰的磁性量子点纳米探针(Mag@QDs-WGA)，并将其引入侧向流分析生物传感器中，构建2种抗体负载检测体系，可在35 min内现场同时检测PA和鼠伤寒沙门氏菌，检测限分别为25 CFU/mL和28 CFU/mL。Hussain等^[83]开发了一种基于免疫磁分离、光散射和机器学习的快速检测PA的芯片平台，免疫磁珠通过抗体-抗原结合特异性捕获目标细菌，样品通过微流控芯片通道获得散射光图，机器学习算法对散射光信号交叉验证评估分类；该方法可以在25 min内特异性和定量检测PA，检测限为1×10² CFU/mL。El Ichi等^[84]以脂多糖(lipopolysaccharide, LPS)抗体为识别元件，设计了一种基于超顺磁性纳米颗粒的微电导免疫传感器，用于无标记且灵敏地检测革兰氏阴性细菌，通过监测细菌与抗体结合后引起的电极表面电导率变化实现对细菌存在的实时检测，结果表明该传感器对PA和鲍曼不动杆菌检出限为10 CFU/mL。

1.4.2 核酸识别元件

核酸识别元件是一种基于序列特异性互补配对的识别机制，通过与目标分子进行互补配对来实现特异性结合的组件。它的关键优势在于其高特异性和高灵敏度，能够通过信号放大实现对低浓度目标分子的检测。目前已结合多种技术应用于PA检测。Tang等^[85]开发了一种基于MNPs的集成化原位PCR方法，利用Fe₃O₄@SiO₂ MNPs吸附并富集DNA后直接进行PCR扩增，同步生成生物素标记的gyrB靶序列；通过探针修饰的MNPs捕获扩增产物，经链霉亲和素-碱性磷酸酶(streptavidin-alkaline phosphatase, SA-ALP)结合生物素后，以化学发光法对PA进行定量检测，检出限低至10 CFU/mL。该方法通过MNPs实现DNA富集、扩增标记与检测一体化，极大地简化了操作流程。Jia等^[86]设计了一种包含可控磁分离和适配体识别的低场磁共振成像(low-field magnetic resonance imaging, LF-MRI)全细胞核酸适配体传感器，即将适配体共价固定在2种不同直径的磁性纳米颗粒(MN10和MN400)上，当PA存在时可与适配体特异性结合形成MN10/细菌/MN400 (MBM)复合物，利用磁场进行磁分离，可将MBM复合物从溶液中快速分离，LF-MRI技术测量剩余溶液的横向弛豫时间(T₂)作为信号读出，通过分析T₂值的变化来定量检测PA；该传感器可在非常低的浓度下检测目标细菌，且无需其他信号增强的策略帮助，整个检测过程约为2 h，检出限为1×10² CFU/mL。

2 总结与展望

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PA作为医院感染病原菌的重要来源之一，严重威胁人类生命健康。加之其高度抗生素耐药特性，使传统治疗变得越发困难，快速且准确的检测手段变得尤为重要。目前，传统的检测方法如平板培养法、PCR法、免疫法等仍未得到有效替代。相反，这些传统方法暴露出了耗时长、操作复杂、成本高等局限性，难以满足快速、灵敏的检测需求。

电化学生物传感器具有低检测限、宽线性范围的优点，优良的材料修饰电极还能放大电化学信号。电化学生物传感器在PA检测中具有广阔的开发前景，有望开发出小型便携式的检测仪器用于现场实际快检。

相比之下，光学通过光信号捕捉目标物同样表现出色。SERS技术检测时长通常较短，远快于传统的培养法，无标记和代谢产物检测则进一步扩展了检测范围。荧光生物检测中，荧光探针和荧光素标记起了主要作用，荧光信号可视化使得结果直观易懂，便于非专业人员理解和操作。SPR技术直接检测分子间的相互作用，不需要对病原菌进行荧光或放射性标记，既能简化流程还能降低检测成本。

CRISPR/Cas12a系统通过结合各类等温核酸扩增技术提高了检测灵敏度，特异性crRNA引导的DNA切割和信号放大，进一步提高了检测效能。MNPs则展现出了优异的磁分离和富集能力，通过与多种检测方法联用，进一步提升了检测灵敏度和准确性。

尽管这些生物传感技术在PA检测中取得了显著进展，但仍面临一些挑战。例如，电化学生物传感器的选择性有待提高；SERS技术的信号稳定性受基底材料和环境干扰的影响；荧光检测的背景干扰和探针稳定性问题仍需解决；SPR仪器设备昂贵，不能有效普及到有限的场地；CRISPR/Cas12a系统在实际样本中的复杂基质干扰也限制了其广泛应用；MNPs的分散性和稳定性问题仍需通过表面功能化修饰来优化。综上所述，针对PA检测的生物传感技术优化任务依旧艰巨。

回顾PA检测技术的发展历程，鲜明体现了从传统方法局限到新型传感技术的突破发展(图1)。早期依赖培养与生化鉴定的传统手段，因耗时久、灵敏度低等问题难以满足即时检测需求；而新兴生物传感技术通过纳米材料、分子识别元件与信号放大策略的协同创新，实现了检测效能的量级提升。

随着纳米技术的不断发展，多学科和多技术交融成为有效推动新型检测技术发展的有力手段，新思维的碰撞通常能推动新型传感检测技术的发展。面向未来，生物传感领域的技术迭代需围绕以下维度展开：(1) 技术融合与材料创新并重，通过开发多功能纳米探针与多模态传感界面，突破单一检测机制的局限；(2) 设备便携化与场景适应性升级，参考技术成熟的小型便携设备开发与智能手机集成技术，推动PA检测从实验室走向临床诊断和现场快检；(3) 复杂样本抗干扰策略优化，设计特异性更强的生物元件，应对复杂基质样品信号干扰问题。通过这些方法优化让PA检测更高效、更精准，在临床诊断、环境监测和食品安全中发挥更加重要的作用。

作者贡献声明

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黄洁洁：撰写文章、文献检索、图表绘制；郑华：提出概念、基金获取、修订完善。

作者利益冲突公开声明

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作者声明不存在任何可能会影响本文所报告工作的已知经济利益或个人关系。

基金

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国家自然科学基金(92360833)

参考文献

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文献

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2025年第65卷第11期

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doi: 10.13343/j.cnki.wsxb.20250327

接收时间：2025-04-21
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出版时间：2025-11-04

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收稿日期：2025-04-21
录用日期：2025-07-01

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National Natural Science Foundation of China(92360833)

国家自然科学基金(92360833)

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广西医科大学生命科学研究院，广西南宁

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Recognition element	Advantages	Disadvantages	References
Antibodies	High specificity, high selectivity, mature technology	Prone to false positives, large molecular weight, poor stability, high cost	[21]
Aptamers	High specificity, good stability, low immunogenicity, scalable in vitro production	Complex screening process, lack of universal systematic evolution of ligand by exponential enrichment (SELEX) standards, requires stability improvement	[22]
Bacteriophages	Strong specificity, low cost, simple operation, protease-insensitive	Immature biosensor integration, requires technical development	[23]
Molecularly imprinted polymers (MIP)	High stability, simple preparation, low cost	Weak binding affinity, low selectivity, challenging large-scale synthesis, susceptible to matrix interference	[24]
Lectins	Broad-spectrum binding, unique glycosyl recognition capability, high stability	Limited selectivity, insufficient affinity and specificity	[25]

Recognition element

Advantages

Disadvantages

References

Antibodies

High specificity, high selectivity, mature technology

Prone to false positives, large molecular weight, poor stability, high cost

[21]

Aptamers

High specificity, good stability, low immunogenicity, scalable in vitro production

Complex screening process, lack of universal systematic evolution of ligand by exponential enrichment (SELEX) standards, requires stability improvement

[22]

Bacteriophages

Strong specificity, low cost, simple operation, protease-insensitive

Immature biosensor integration, requires technical development

[23]

Molecularly imprinted polymers (MIP)

High stability, simple preparation, low cost

Weak binding affinity, low selectivity, challenging large-scale synthesis, susceptible to matrix interference

[24]

Lectins

Broad-spectrum binding, unique glycosyl recognition capability, high stability

Limited selectivity, insufficient affinity and specificity

[25]

Sensor type	Electrode	Functional material system	Recognition element	Detection sample	Limit of detection (CFU/mL)	Linear range (CFU/mL)	References
Amperometric	Screen-printed three-electrode system (SPE)	Gold nanoparticles (GNPs)	Aptamer	Water	60	6.0×10¹-6.0×10⁷	[28]
Gold electrode	AuNPs/Super P/Cu-ZrMOF	Antibody, aptamer	Urine	2	10¹-10⁷	[29]
Carbon screen-printed electrode (CSPE)	MIL-101(Cr)/MWCNT/AgNPs/c-g-C₃N₄	Aptamer	Serum	1	10¹-10⁷	[30]
GCE	AuNPs	Aptamer-MIP	Serum	1	10¹-10⁷	[31]
GCE	CCLP/AuNPs	Monoclonal antibody	Water	9×10²	10¹-10⁷	[32]
GCE	HZIFs-8/Fc-GO	Aptamer	Urine	1	1.2×10¹-1.2×10⁷	[33]
Au-SPEs	MB	Aptamer	Water	8	0-10¹⁰	[34]
GCE	CoFe₂O₄/AgNPs	Pyocyanin	Apple, beef, water, chicken, fish, egg, soil, milk	4.0×10^-3	1-23	[35]
Impedimetric	GCE	AgNPs	Aptamer	Serum	33	10²-10⁷	[36]
GCE	NCs/Glu	Aptamer	Serum	3	10¹-10⁷	[37]
GCE	CNCNF	Aptamer	Serum	1	10¹-10⁷	[38]
ECL	GCE	Carboxyl graphene	Phage	Milk, glucose injection, urine	56	1.4×10²-1.4×10⁶	[39]
Piezoelectric	Gold (Au) interdigital electrode (Au IDE)	Polyadenylated DNA/MB	Aptamer	Buﬀer, blood	9 (buﬀer), 52 (blood)	8.1×10¹-8.1×10⁵ (buﬀer), 1.9×10²-1.0×10⁶ (blood)	[40]

Sensor type

Electrode

Functional material system

Recognition element

Detection sample

Limit of detection (CFU/mL)

Linear range (CFU/mL)

References

Amperometric

Screen-printed three-electrode system (SPE)

Gold nanoparticles (GNPs)

Aptamer

Water

6.0×10¹-6.0×10⁷

[28]

Gold electrode

AuNPs/Super P/Cu-ZrMOF

Antibody, aptamer

Urine

10¹-10⁷

[29]

Carbon screen-printed electrode (CSPE)

MIL-101(Cr)/MWCNT/AgNPs/c-g-C₃N₄

Aptamer

Serum

10¹-10⁷

[30]

GCE

AuNPs

Aptamer-MIP

Serum

10¹-10⁷

[31]

GCE

CCLP/AuNPs

Monoclonal antibody

Water

9×10²

10¹-10⁷

[32]

GCE

HZIFs-8/Fc-GO

Aptamer

Urine

1.2×10¹-1.2×10⁷

[33]

Au-SPEs

Aptamer

Water

0-10¹⁰

[34]

GCE

CoFe₂O₄/AgNPs

Pyocyanin

Apple, beef, water, chicken, fish, egg, soil, milk

4.0×10^-3

1-23

[35]

Impedimetric

GCE

AgNPs

Aptamer

Serum

10²-10⁷

[36]

GCE

NCs/Glu

Aptamer

Serum

10¹-10⁷

[37]

GCE

CNCNF

Aptamer

Serum

10¹-10⁷

[38]

ECL

GCE

Carboxyl graphene

Phage

Milk, glucose injection, urine

1.4×10²-1.4×10⁶

[39]

Piezoelectric

Gold (Au) interdigital electrode (Au IDE)

Polyadenylated DNA/MB

Aptamer

Buﬀer, blood

9 (buﬀer), 52 (blood)

8.1×10¹-8.1×10⁵ (buﬀer), 1.9×10²-1.0×10⁶ (blood)

[40]

Sensor type	Signal system (amplifier/ reporter/quencher)	Recognition element	Detection sample	Limit of detection (CFU/mL)	Linear range (CFU/mL)	References
SERS	AuNPs (amplifier), 4-MBA (reporter)	Aptamer	Water, chicken meat	20	10²-10⁷	[49]
Fe₃O₄@Au (amplifier), DTNB (reporter)	Cecropin 1, antibody	Urine	12 cell/mL	/	[50]
AuNSs (amplifier), DTNB (reporter)	WGA, antibody	Urine	5	10¹-10⁶	[51]
AuNPs (amplifier), DTNB (reporter)	Single-guide RNA, CRISPR/dCas9	Urine	1	1-10⁶	[52]
Au (amplifier)	/	Blood	5×10³	/	[53]
Fluorescent	TPE (reporter)	Aptamer	Water, orange juice, milk	1×10²	10²-10⁸	[54]
CDs (reporter), GO (quencher)	Aptamer	Water	9	10¹-10⁷	[55]
PDA-PEI copolymer dots (reporter)	Dual aptamer	Milk, orange juice, popsicle	1	10¹-10⁷	[56]
HCR (amplifier), FAM (reporter), BHQ-1 (quencher)	Aptamer	Urine	37	10²-10⁷	[57]
FAM (reporter)	DNAzyme (PAE-1)	Water, pekoe tea, peach juice, hawthorn juice	1.2	/	[58]
FAM-cDNA (reporter), GOQDs (quencher)	Aptamer	Water, orange juice, popsicle	1×10²	1.28×10³-2.0×10⁷	[59]
SPR	MG-NPA (amplifier)	Aptamer	/	30 CFU/assay	/	[60]
Au nanotriangle array (amplifier)	Aptamer	/	10	10¹-10³	[61]

Sensor type

Signal system (amplifier/ reporter/quencher)

Recognition element

Detection sample

Limit of detection (CFU/mL)

Linear range (CFU/mL)

References

SERS

AuNPs (amplifier), 4-MBA (reporter)

Aptamer

Water, chicken meat

10²-10⁷