免疫分析方法在氧氟沙星残留检测中的研究进展

doi:10.16178/j.issn.0528-9017.20240489

摘要/Abstract

摘要：

氧氟沙星(ofloxacin, OFLX)是一种主要的氟喹诺酮类(FQs)药物,因其抗菌性好、价格低廉而常被用于人类和动物的医疗保健中,但其残留物对人类健康造成的风险巨大,引起人们的高度重视。鉴于OFLX在食品和环境中有较多残留,建立对OFLX的即时检测方法势在必行。与传统的色谱、光谱检测技术相比,免疫分析法在特异性、检测效率、现场适用性和前处理的简便性等方面具有突出优势。该文综述了OFLX抗体的制备和免疫分析技术在OFLX残留检测中的应用现状,比较了酶联免疫法、免疫层析法、荧光免疫法、免疫传感器法、生物条形码免疫分析法、核酸适配体技术等方法的优缺点,并对OFLX免疫检测方法的发展趋势进行了展望,以期为该抗生素快速检测方法的建立和应用提供参考。

关键词: 氧氟沙星, 抗体, 免疫分析, 生物传感器, 核酸适配体

Abstract:

Ofloxacin (OFLX), a major kind of fluoroquinolones(FQs), is commonly used in human and animal healthcare due to its antimicrobial properties and low cost. However, the risk to human health posed by its residues has attracted considerable attention. In view of the residues of OFLX in the food and environment, it is imperative to establish its immediate detection. Compared with traditional chromatographic and spectroscopic detection techniques, immunoassay has outstanding advantages in terms of specificity, detection efficiency, field applicability and simplicity of pre-treatment. This paper reviewed the preparation of OFLX antibodies and the current application of immunoassay techniques in OFLX residue detection, compared the advantages and disadvantages of enzyme-linked immunoassay, immunochromatographic method, fluorescence immunoassay, immunosensor, bio-barcode immunoassay, and nucleic acid aptamer, etc., and looked forward to the development trend of the immunoassay methods of OFLX with the aim of providing a reference for the establishment and application of rapid antibiotic detection methods.

Key words: ofloxacin, antibody, immunoassay, biosensor, nucleic acid aptamer

中图分类号:

S859.79+6

杨兴东, 曲扬, 胡骁飞, 李俐俐. 免疫分析方法在氧氟沙星残留检测中的研究进展[J]. 浙江农业科学, 2025, 66(3): 742-749.

YANG Xingdong, QU Yang, HU Xiaofei, LI Lili. Research progress of immunoassay methods in the detection of ofloxacin residues[J]. Journal of Zhejiang Agricultural Sciences, 2025, 66(3): 742-749.

图/表 6

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合成免疫原的方法	免疫动物	抗体类型	半数抑制浓度(IC₅₀)/ (ng·mL^-1)	检测限/ (ng·mL^-1)	参考文献
碳二亚胺法	BALB/c小鼠	mAb	14.580	1.590	[17]
NHS酯法	新西兰白兔	pAb	0.500 (R, S-OFLX)	0.300~0.400	[18]
			3.100 (S-OFLX)
碳二亚胺法	BALB/c小鼠	mAb	0.696	0.200	[19]
碳二亚胺法	BALB/c小鼠	mAb	7.000	0.500	[20]
碳二亚胺法	BALB/c小鼠	pAb	19.970	0.240	[21]
碳二亚胺法	BALB/c小鼠	mAb	12.430 (R-OFLX) 0.650 (S-OFLX)	—	[22]
NHS酯法	新西兰白兔	pAb	0.430 (R-OFLX) 0.300(S-OFLX)	—	[23]
碳二亚胺法	BALB/c小鼠	mAb	0.893	0.151	[24]
NHS酯法	新西兰白兔	pAb	1.840	—	[25]

合成免疫原的方法	免疫动物	抗体类型	半数抑制浓度(IC₅₀)/ (ng·mL^-1)	检测限/ (ng·mL^-1)	参考文献
碳二亚胺法	BALB/c小鼠	mAb	14.580	1.590	[17]
NHS酯法	新西兰白兔	pAb	0.500 (R, S-OFLX)	0.300~0.400	[18]
			3.100 (S-OFLX)
碳二亚胺法	BALB/c小鼠	mAb	0.696	0.200	[19]
碳二亚胺法	BALB/c小鼠	mAb	7.000	0.500	[20]
碳二亚胺法	BALB/c小鼠	pAb	19.970	0.240	[21]
碳二亚胺法	BALB/c小鼠	mAb	12.430 (R-OFLX) 0.650 (S-OFLX)	—	[22]
NHS酯法	新西兰白兔	pAb	0.430 (R-OFLX) 0.300(S-OFLX)	—	[23]
碳二亚胺法	BALB/c小鼠	mAb	0.893	0.151	[24]
NHS酯法	新西兰白兔	pAb	1.840	—	[25]

方法	优点	缺点	特点
酶联免疫法	前处理简单、成本较低、设备投资少、应用广泛	重复性较差,干扰因素多,易出现假阳性结果	酶标记,酶促反应
荧光免疫法	操作简便,灵敏度较高、安全	易受背景干扰、仪器要求高、结果不便长期保存	荧光标记
胶体金免疫分析法	无需前处理、现场快速筛选、结果易判定	肉眼观察颜色变化只能定性判定,定量检测需用读条仪	胶体金标记
免疫传感器法	选择性好,灵敏度较高、快速、自动化检测	制备复杂、难度大,检测元件要求高	传感器信号的放大和识别
生物条形码免疫分析法	操作简便、灵敏度高、结果准确	制备过程困难,所需设备昂贵	多重信号放大,抗体为特异性识别元件
核酸适配体技术	亲和力和特异性强,易于制备和修饰,应用范围广	筛选过程繁杂,成功率不高	与靶分子的特异性结合

方法	优点	缺点	特点
酶联免疫法	前处理简单、成本较低、设备投资少、应用广泛	重复性较差,干扰因素多,易出现假阳性结果	酶标记,酶促反应
荧光免疫法	操作简便,灵敏度较高、安全	易受背景干扰、仪器要求高、结果不便长期保存	荧光标记
胶体金免疫分析法	无需前处理、现场快速筛选、结果易判定	肉眼观察颜色变化只能定性判定,定量检测需用读条仪	胶体金标记
免疫传感器法	选择性好,灵敏度较高、快速、自动化检测	制备复杂、难度大,检测元件要求高	传感器信号的放大和识别
生物条形码免疫分析法	操作简便、灵敏度高、结果准确	制备过程困难,所需设备昂贵	多重信号放大,抗体为特异性识别元件
核酸适配体技术	亲和力和特异性强,易于制备和修饰,应用范围广	筛选过程繁杂,成功率不高	与靶分子的特异性结合

检测类型	材料	基质	线性范围/ (ng·mL^-1)	检测限/ (ng·mL^-1)	参考文献
电化学	PPy-Au nanocluster	PBS	0.08~410.00	0.03	[35]
电化学	multi-enzyme-AuNFs	HEPES	0.26~25.60(S-OFL)	0.15	[36]
			0.37~12.80 (R-OFL)	0.30
光学	UCNPs (β-NaYF4/Yb3+/Er3+)	PBS	—	3.61	[37]
光学	nanoMIPs (APTMS)	河水牛奶	—	57.46(河水) 46.98(牛奶)	[38]
压电	MWCNTs	牛奶	30.0~650.0	9.00	[39]