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基于链霉亲和素-生物素系统和 O-羧甲基壳聚糖靶标钆探针的核磁共振生物传感器快速检测牛奶中的沙门氏菌。

Rapid detection of Salmonella in milk by a nuclear magnetic resonance biosensor based on the streptavidin-biotin system and O-carboxymethyl chitosan target gadolinium probe.

机构信息

State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China.

Key Laboratory of Bioprocess Engineering of Jiangxi Province, Jiangxi Science and Technology Normal University, Nanchang 330013, China.

出版信息

J Dairy Sci. 2021 Nov;104(11):11486-11498. doi: 10.3168/jds.2021-20716. Epub 2021 Aug 26.

DOI:10.3168/jds.2021-20716
PMID:34454766
Abstract

Rapid and sensitive detection of foodborne pathogens is of great importance for food safety. Here, a set of nuclear magnetic resonance (NMR) biosensors based on a O-carboxymethyl chitosan target gadolinium (Gd) probe was developed to quickly detect Salmonella in milk by combining NMR technology and bioimmunotechnology with membrane filtration technology. First, O-carboxymethyl chitosan (O-CMC) was biotinylated to prepare biotinylated O-carboxymethyl chitosan (biotin-O-CMC) through amide reaction, and biotinylated magnetic complexes (biotin-O-CMC-Gd) were obtained by using O-CMC, which has strong chelating adsorption on Gd. The target probe was obtained by combining biotin-O-CMC-Gd with the biotinylated antibody (biotin-antibody) via streptavidin (SA) by introducing the SA-biotin system. Then, Salmonella was captured by the target probe through antigen-antibody interaction. Finally, NMR was used to measure the longitudinal relaxation time (T) of the filtrate collected by membrane filtration. This NMR biosensor with good specificity and high efficiency can detect Salmonella with the sensitivity of 1.8 × 10 cfu/mL within 2 h; in addition, it can realize the detection of complex samples because of its strong anti-interference capability and may open up a new method for rapid detection of Salmonella, which has a great application potential.

摘要

快速灵敏地检测食源性致病菌对于食品安全至关重要。本研究结合膜过滤技术、生物免疫技术和核磁共振(NMR)技术,利用一种基于壳聚糖羧甲基(O-CMC)靶钆(Gd)探针的 NMR 生物传感器快速检测牛奶中的沙门氏菌。首先,通过酰胺反应将壳聚糖羧甲基化以制备生物素化 O-CMC(biotin-O-CMC),然后利用 O-CMC 对 Gd 具有很强的螯合吸附作用,得到生物素化磁复合物(biotin-O-CMC-Gd)。通过引入链霉亲和素(SA)-生物素系统,将 biotin-O-CMC-Gd 与生物素化抗体(biotin-antibody)结合,制备得到靶探针。然后,通过抗原抗体相互作用,靶探针捕获沙门氏菌。最后,通过膜过滤收集滤液并利用 NMR 测量其纵向弛豫时间(T)。该 NMR 生物传感器具有良好的特异性和高效率,可在 2 h 内检测到灵敏度为 1.8×10 cfu/mL 的沙门氏菌;此外,由于其具有较强的抗干扰能力,可实现对复杂样品的检测,为沙门氏菌的快速检测开辟了新途径,具有广阔的应用前景。

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