基于适配体的光学和电化学法对食源性致病菌检测的研究进展

doi:10.3864/j.issn.0578-1752.2021.11.014

Abstract

Abstract:

Food safety has become a heated topic attracting widespread attention from the society due to the diversity of food and complexity of food production system. Food-borne diseases caused by microorganisms have the highest rates in food safety problem. The detection of food-borne pathogenic bacteria is the key link for the food-borne disease prevention and control. The plate counting method is rated as the gold standard for microbial detection, but the signal amplification was achieved by the growth of individual bacterial cells into visible colony during the detection of pathogenic bacteria, so the detection time is longer (3-7 days). Although polymerase chain reaction (PCR) and enzyme-linked immunoabsorbent assay (ELISA) are now applied in the detection of food-borne pathogenic bacteria, they are not suitable for timely and rapid on-site detection due to time-consuming pretreatment, complex operations and false positive results. Aptamers (Apt) are oligonucleotides that are isolated from combinatorial DNA library via systemic evolution of ligands by exponential enrichment (SELEX) technology, which present great potential in the field of detection of toxin, heavy metal, antibiotics, food pathogens and other small molecules due to their small size, easy synthesis and modification. Currently, many researchers of domestic and overseas have developed various detection methods by using aptamer as bio-recognition elements. This paper reviewed common food-borne pathogenic bacteria, traditional detection methods of food-borne pathogenic bacteria, and the electrochemical and optical aptasensors for detection of food-borne pathogenic bacteria in recent years. The descriptions covered the detection strategy of each method and provided details such as the detection time, range and limit. Finally, the paper pointed out the flaw of aptasensor in food safety detection, and the research development tendency was prospected, which provided bases for the further related work.

Key words: food-born pathogen bacteria, aptamer, rapid detection, food safety

HUI YuanYuan,PENG HaiShuai,WANG BiNi,ZHANG FuXin,LIU YuFang,JIA Rong,REN Rong. Research Progress of Food-Borne Pathogen Detection Based on Electrochemical and Optical Aptasensors[J].Scientia Agricultura Sinica, 2021, 54(11): 2419-2433.

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URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2021.11.014

https://www.chinaagrisci.com/EN/Y2021/V54/I11/2419

Figures/Tables 3

Table 1

Table 2

Aptamers of food-borne pathogens"

食源性致病菌 Pathogen	菌型 Bacterial type	筛选方法 Screening method	靶标 Target	适配体序列 Sequence (5′-3′)	参考文献 Reference
大肠杆菌 Escherichia coli	O157:H7	Non-SELEX	脂多糖 LPS	TATGGCGTGGCAAGCTTGGCCCGCTTCTCAAGCATGGTTATCTAC	[42]
		SELEX	全菌 Whole bacteria	ACCAGTAGACTTTCAACTTTACTGCCATCGTGTGCCCTAA	[43]
		Non-SELEX	脂多糖 LPS	CCGGACGCTTATGCCTTGCCATCTACAGAGCAGGTGTGACGG	[44]
	EPEC	SELEX	全菌 Whole bacteria	ACAGTGCTCGGGATATATCAATATGTCACCTCGGCTAATG	[45]
	O55:B5	Non-SELEX	脂多糖 LPS	TAGCCGGATCGCGCTGGCCAGATGATATAAAGGGTCAGCCCCCCA	[46]
沙门氏菌 Salmonella	鼠伤寒 Typhimurium	Whole-cell-SELEX	全菌 Whole bacteria	ACGGGCGTGGGGGCAATGCCTGCTTGTAGGCTTCCCCTGTGCGCG	[47]
		SELEX	全菌 Whole bacteria	TATGGCGGCGTCACCCGACGGGGACTTGACATTATGACAG	[48]
		SELEX	IVB型菌毛 IVB pili	GGGAACAGUCCGAGCCUCACUGUUAUCCGAUAGCAGCGCGGGAUGAGGGUCAAUGCGUCAUAGGAUCCCGC	[49]
	O8	Whole-cell-SELEX	全菌 Whole bacteria	GATCCGGGCCTCATGTCGAACACCCCCCAACTAAAACAACAAAACACCACCGCCATTGAGCGTTTATTCTGAGCTCCCA	[50]
	O8	SELEX	全菌 Whole bacteria	TGATCCGGGCCTCATGTCGAACCCACACCCCACAACCACCCAGCCCCAGCCCGCTATTGAGCGTTTATTCTGAGCTCCCA	[51]
金黄色葡萄球菌 Staphylococcus aureus		SELEX	磷壁酸 Teichoic acid	GGGAGUUUUGAUACGGCUUCAUGCAGUAAUGUUUUUAU	[52]
		SELEX	肠毒素C1 Enterotoxin C1	AGCAGCACAGAGGTCAGATGTATACTTCTAAAATTTGTTTGTATCTACGATGTTCTTCGTCCTATGCGTGCTACCGTGAA	[53]
		SELEX	肠毒素B Enterotoxin B	AGCAGCACAGAGGTCAGATGTATACTTCTAAAATTTGTTTGTATCTACGATGTTCTTCGTCCTATGCGTGCTACCGTGAA	[54]
		SELEX	肠毒素B Enterotoxin B	TGCAGGATCCGGTATCCGTGGACGGTGTGCAGGATCCGGTATCCGTGGGCACGAGAATTCCTCCGTTGCG	[55]
		SELEX	肠毒素A Enterotoxin A	TACTTATGCATTTCCTCCCACGATCTTATTTGAGAGTGAC	[56]
单核增生李斯特菌 Listeria monocytogenes		Whole-cell-SELEX	全菌 Whole bacteria	TGGGAGCTCAGAATAAACGCTCAACTTTGTTCTTCTTTGCTTTTTTTTTCTTTTTTTGTTCGACATGAGGCCCGGATCA	[57]
阪崎肠杆菌 Cronobacter sakazakii		Whole-cell-SELEX	全菌 Whole bacteria	ATAGGAGTCACGACGACCAGGCCGCCCGGGGACGGGGGCGGCGGGGAGGAGGGCGGCGGGTATGTGCGTCTACCTCTTGA	[58]
蜡样芽孢杆菌 Bacillus cereus		SELEX	芽孢 Spore	CATCCGTCACACCTGCTCGGTGCAGACCCATAGGGGGGGCGTGCGGATGTAGGAGTAGGGTGTTGGCTCCCGTATC	[59]
铜绿假单胞菌 Pseudomonas aeruginosa		Whole-cell-SELEX	全菌 Whole bacteria	CCCCCGTTGCTTTCGCTTTTCCTTTCGCTTTTGTTCGTTTCGTCCCTGC TTCCTTTCTTG	[60]
副溶血性弧菌 Vibrio parahemolyticus		Whole-cell-SELEX	全菌 Whole bacteria	ATAGGAGTCACGACGACCAGAATCTAAAAATGGGCAAAGAAACAGTGACTCGTTGAGATACTTATGTGCGTCTACCTCTTGACTAAT	[61]

Table 2

Table 3

Comparison of analytical properties of the aptasensors for food-borne pathogen detection"

传感器类型 Sensor type	方法 Method	策略 Strategy	检测范围 Detection range (cfu·mL^-1)	检测限 LOD (cfu·mL^-1)	参考文献 Reference
电化学 Electrochemical	将适配体固定在电极表面,靶标与适配体的结合导致电极表面阻抗增大,通过检测电化学信号的变化检测靶标 The aptamer is modified on the surface of the electrode, and the combination of the target and the aptamer leads to an increase in the surface impedance of the electrode. The target is detected by detecting the change of electrochemical signal	rGO-TiO2复合纳米材料信号放大 RGO -TiO2 composite nanomaterial signal amplification	10¹—10⁸	10	[62]
		rGO-CNT复合纳米材料信号放大 rGO-CNT composite nanomaterial signal amplification	10¹—10⁸	10	[63]
		聚[吡咯-co-3-羧基吡咯]共聚物信号放大 poly[pyrrole-co-3-carboxyl-pyrrole]copolymer signal amplification	10²—10⁸	3	[64]
	三明治法 Sandwich assay	适配体-靶标-MB@MI纳米材料 Apt-target-MB@MI	10²—10⁸	32	[65]
	三明治法 Sandwich assay	抗体/磁性纳米颗粒-靶标-适配/脲酶复合物 Antibody/magnetic nanoparticles - target - aptamer/urease complex	10¹—10⁴	10	[66]
	三明治法 Sandwich assay	滚环扩增 Rolling cycle amplification	2.2—2.2×10⁸	2	[67]
比色 Colorimetric	NaCl-AuNPs	高浓度的盐会引起AuNPs聚集,颜色从酒红色变为蓝色 high-salt induce AuNPs aggregation from red to blue	_	10⁵	[68]
			10²—10⁷	56	[69]
			10⁵—10⁸	7.2×10⁵	[70]
	过氧化氢氧化法 H₂O₂ oxidation	7-四链体-heminDNA酶可催化四甲基联苯胺（TMB）-过氧化氢（H₂O₂）体系发生反应,颜色从无色变为蓝色 G4/Hemin DNAzyme can catalyze TMB-H₂O₂ reaction system, and colour of the substrates changing from colourless to blue	10²—10⁷	10	[71]
		纳米模拟酶可催化TMB-H₂O₂体系发生反应,颜色从无色变为蓝色 Nanozymes can catalyze TMB-H₂O₂ reaction system, and colour of the substrates changing from colourless to blue	11—1.1×10⁵	11	[72]
			10²—10⁶	10	[73]
荧光 Fluorescent	荧光的产生 Fluorescence generation	TPE-OH 能在牛血清白蛋白（BSA）微球中聚集,产生明亮的荧光信号 TPE-OH aggregates in bovine serum albumin (BSA) microspheres producing bright fluorescent signal	10¹—10⁶	10	[74]
	荧光的猝灭 Fluorescence quenching	MoS₂纳米材料对于荧光素（FAM）的猝灭 Quenching of MoS₂ nanomaterials to fluorescein (FAM)	10²—10⁵	10	[75]
	荧光的猝灭 Fluorescence quenching	CIP@MWCNT纳米材料对于荧光素（FAM）的猝灭 Quenching of CIP@MWCNT nanomaterials to fluorescein (FAM)	7.15—10³	3.15×10²	[76]
	荧光染料AccuBlue? Fluorescent dye AccuBlue?	靶标与适配体的结合会改变荧光染料的荧光信号 The binding of target and aptamer changed the fluorescence signal of fluorescent dye	50—10⁶	25	[77]
	荧光染料罗丹明B Fluorescent dye Rhodamine B		1530—96938	464	[78]
化学发光 Chemiluminescence	化学发光共振能量转移 Chemiluminescence resonance energy transfer	Co²⁺增强/ABEI功能化花状纳米金和滚环扩增技术 Co²⁺ enhanced /ABEI functionalized flower-like nanogold and roll ring amplification techniques	32—3.2 ×10⁶	10	[79]
化学发光 Chemiluminescence	化学发光共振能量转移 Chemiluminescence resonance energy transfer		50—1.5×10⁵	15	[80]
表面增强拉曼散射 SERS	将适配体修饰在SERS基地上,当靶标与适配体结合后会产生拉曼信号变化,通过对拉曼变化信号的检测进而检测靶标菌 The aptamer is modified on the SERS base. When the target and the aptamer are combined, the Raman signal will change, and the target bacteria can be detected by detecting the Raman change signal	适配体和罗丹明 B 同时修饰的金纳米棒作为拉曼信号探针 The gold nanorods modified by aptamer and rhodamine B was used as Raman signal probe	10¹—10⁴	3	[81]
		适配体和拉曼分子修饰的金纳米颗粒作为拉曼信号探针 The nanoparticles modified by aptamer and raman molecules was used as Raman signal probe	10²—10⁷	35/15	[82]
		适配体和罗丹明修饰的Au@Ag 核壳纳米材料作为拉曼信号探针 The Au@Ag core-shell nanomaterials modified by ROX and aptamer was used as Raman signal probe	15—1.5×10⁵	15	[83]
表面等离子体共振 Surface plasmon resonance	将适配体修饰在等离子共振装置表面上。当靶标菌存在时共振装置折射率、反射光等会发生变化,通过对光变化信号的检测可以检测靶标 The aptamer is modified on the surface of the plasmon resonance device. When the target bacteria are present, the refractive index and reflected light of the resonance device will change, and the target can be detected by detecting the light change signal	使用Ω形光纤探针 Using a novel Ω-shaped ?ber-optic probe	5.0×10²— 1.0×10⁸	5	[84]

Table 3

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传统方法 Method	优势 Advantage	缺陷 Limitation
培养法 Culture method	检测限低、可靠性强 Low detection limit and high credibility	耗时、耗力 Time-consuming and laborious
基于免疫的方法 Immunology-based method	准确度高 High accuracy	灵敏度低 Low sensitivity
基于PCR的方法 PCR-based method	灵敏度高 High sensitivity	假阴性 False negative

Research Progress of Food-Borne Pathogen Detection Based on Electrochemical and Optical Aptasensors

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