功能核酸用于致病菌检测的研究进展

doi:10.19586/j.2095-2341.2022.0089

生物技术进展 ›› 2023, Vol. 13 ›› Issue (1): 30-38.DOI: 10.19586/j.2095-2341.2022.0089

功能核酸用于致病菌检测的研究进展

赵文卓(), 李成勋, 胡作建, 余红秀()

复旦大学生物医学研究院&附属口腔医院，上海 200032

收稿日期:2022-05-26 接受日期:2022-08-18 出版日期:2023-01-25 发布日期:2023-02-07
通讯作者: 余红秀
作者简介:赵文卓 E-mail: 21211510023@m.fudan.edu.cn；
基金资助:
国家重点研发计划项目(2021YFC2701802);上海市口腔医院科技创新人才培养计划(SSH-2022-KJCX-A01)

Research Progress of Functional Nucleic Acid Used in Pathogenic Bacteria Detection

Wenzhuo ZHAO(), Chengxun LI, Zuojian HU, Hongxiu YU()

Institutes of Biomedical Sciences & Shanghai Stomatological Hospital，Fudan University，Shanghai 200032，China

Received:2022-05-26 Accepted:2022-08-18 Online:2023-01-25 Published:2023-02-07
Contact: Hongxiu YU

摘要/Abstract

摘要：

由食源性致病菌引发的疾病对人类健康构成巨大威胁。虽然一些致病菌如金黄色葡萄球菌、大肠杆菌和沙门氏菌等在诊断和预防方面已经取得了重大进展，但开发快速、高效、低成本的检测方法仍然是一项挑战。功能核酸（functional nucleic acids，FNAs）是一类功能超出核酸常规遗传作用的核酸，主要包括天然的核酶（RNAzymes）、核糖开关（riboswitches）以及体外通过指数富集配体系统进化技术（systematic evolution of ligands by exponential enrichment，SELEX）筛选的适配体（aptamers）、核酶（RNAzymes）和脱氧核酶（DNAzymes）。适配体和脱氧核酶因具有较高的稳定性、特异性和可设计性，使其成为病原微生物识别的理想工具，近年来在生物传感和医学诊断领域备受关注。综述了功能核酸的筛选原理和流程、适配体及具有RNA裂解活性的脱氧核酶（RNA cleavage deoxyribozymes，RCDs）在致病菌检测中的应用进展和面临的挑战，并对其未来的发展前景进行了展望。

关键词: 功能核酸, SELEX技术, 致病菌, 生物传感器

Abstract:

Diseases caused by food-borne pathogenic bacteria pose a great threat to human health. Although some pathogenic bacteria such as Staphylococcus aureus， Escherichia coli and Salmonella enterica have made great progress in diagnosis and prevention， it is still a challenge to develop fast， efficient and low-cost detection methods. Functional nucleic acids （FNAs） are a kind of nucleic acids with functions beyond the conventional genetic function of nucleic acids， mainly including natural ribozymes， riboswitches， as well as aptamers， RNAzymes and deoxyribozymes screened by exponential enrichment ligand system evolution （SELEX） in vitro. Because of its high stability， specificity and designability， aptamers and DNAzymes have become an ideal tool for identifying pathogenic microorganisms， which has attracted much attention in the fields of biosensing and medical diagnosis in recent years. This paper reviewed the screening principle and process of functional nucleic acids， the application progress and challenges of aptamers and RNA cleavage deoxyribozymes （RCDs） in the detection of pathogenic bacteria， and prospected their future development prospects.

Key words: functional nucleic acids, SELEX technology, pathogenic bacteria, biosensor

中图分类号:

R440

赵文卓, 李成勋, 胡作建, 余红秀. 功能核酸用于致病菌检测的研究进展[J]. 生物技术进展, 2023, 13(1): 30-38.

Wenzhuo ZHAO, Chengxun LI, Zuojian HU, Hongxiu YU. Research Progress of Functional Nucleic Acid Used in Pathogenic Bacteria Detection[J]. Current Biotechnology, 2023, 13(1): 30-38.

图/表 6

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检测方法	适用场所	优缺点	检测对象	检测元件	检测限/（CFU·mL^-1）	参考文献
荧光法	实验室、机构检测	检出限较低，但需要相关仪器设备，成本高	苏云金芽孢杆菌（Bacillus thuringiensis）	适配体	10³	[23]
			副溶血弧菌和鼠伤寒沙门氏菌（Vibrio parahaemolyticus and Salmonella typhimurium）	适配体	5×10³	[24]
			大肠杆菌ATCC 8739（Escherichia coli）	适配体	3	[25]
			鼠伤寒沙门氏菌（Salmonella typhimurium）	适配体	50	[26]
			大肠杆菌（Escherichia coli）	RCDs	10³	[27]
			肺炎克雷伯菌（Klebsiella Pneumoniae）	RCDs	10⁵	[28]
			鳗弧菌（Vibrio anguillarum）	RCDs	4×10³	[29]
比色法	现场检测、实验室、机构检测	直观快速，不需要仪器支撑，成本低，但结果不易量化	金黄色葡萄球菌（Staphylococcus aureus）	适配体	10⁸	[30]
			阪崎肠杆菌（Cronobacter sakazakii）	适配体	7.1×10³	[31]
			幽门螺旋杆菌（Helicobacter pylori）	RCDs	10⁴	[32]
			金黄色葡萄球菌（Staphylococcus aureus）	RCDs	10⁵	[33]
电化学法	现场检测、实验室、机构检测	灵敏度高，结果直观，但需要相关电子设备，成本较高	金黄色葡萄球菌（Staphylococcus aureus）	适配体	41	[34]
			金黄色葡萄球菌（Staphylococcus aureus）	适配体	39（缓冲液） 414（自来水）	[35]
			大肠杆菌（Escherichia coli）	RCDs	10³	[36]

检测方法	适用场所	优缺点	检测对象	检测元件	检测限/（CFU·mL^-1）	参考文献
荧光法	实验室、机构检测	检出限较低，但需要相关仪器设备，成本高	苏云金芽孢杆菌（Bacillus thuringiensis）	适配体	10³	[23]
			副溶血弧菌和鼠伤寒沙门氏菌（Vibrio parahaemolyticus and Salmonella typhimurium）	适配体	5×10³	[24]
			大肠杆菌ATCC 8739（Escherichia coli）	适配体	3	[25]
			鼠伤寒沙门氏菌（Salmonella typhimurium）	适配体	50	[26]
			大肠杆菌（Escherichia coli）	RCDs	10³	[27]
			肺炎克雷伯菌（Klebsiella Pneumoniae）	RCDs	10⁵	[28]
			鳗弧菌（Vibrio anguillarum）	RCDs	4×10³	[29]
比色法	现场检测、实验室、机构检测	直观快速，不需要仪器支撑，成本低，但结果不易量化	金黄色葡萄球菌（Staphylococcus aureus）	适配体	10⁸	[30]
			阪崎肠杆菌（Cronobacter sakazakii）	适配体	7.1×10³	[31]
			幽门螺旋杆菌（Helicobacter pylori）	RCDs	10⁴	[32]
			金黄色葡萄球菌（Staphylococcus aureus）	RCDs	10⁵	[33]
电化学法	现场检测、实验室、机构检测	灵敏度高，结果直观，但需要相关电子设备，成本较高	金黄色葡萄球菌（Staphylococcus aureus）	适配体	41	[34]
			金黄色葡萄球菌（Staphylococcus aureus）	适配体	39（缓冲液） 414（自来水）	[35]
			大肠杆菌（Escherichia coli）	RCDs	10³	[36]

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功能核酸用于致病菌检测的研究进展

Research Progress of Functional Nucleic Acid Used in Pathogenic Bacteria Detection

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