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一种利用量子点和结合不同多克隆抗体的磁珠快速检测布鲁氏菌的方法。

A Rapid Detection Method of Brucella with Quantum Dots and Magnetic Beads Conjugated with Different Polyclonal Antibodies.

作者信息

Song Dandan, Qu Xiaofeng, Liu Yushen, Li Li, Yin Dehui, Li Juan, Xu Kun, Xie Renguo, Zhai Yue, Zhang Huiwen, Bao Hao, Zhao Chao, Wang Juan, Song Xiuling, Song Wenzhi

机构信息

Department of Health Laboratory, School of Public Health, Jilin University, 130021, Changchun, China.

School of Public Health, Xuzhou Medical University, 221000, Xuzhou, China.

出版信息

Nanoscale Res Lett. 2017 Dec;12(1):179. doi: 10.1186/s11671-017-1941-z. Epub 2017 Mar 9.

Abstract

Brucella spp. are facultative intracellular bacteria that cause zoonotic disease of brucellosis worldwide. Traditional methods for detection of Brucella spp. take 48-72 h that does not meet the need of rapid detection. Herein, a new rapid detection method of Brucella was developed based on polyclonal antibody-conjugating quantum dots and antibody-modified magnetic beads. First, polyclonal antibodies IgG and IgY were prepared and then the antibody conjugated with quantum dots (QDs) and immunomagnetic beads (IMB), respectively, which were activated by N-(3-dimethylaminopropyl)-N'-ethylcar-bodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) to form probes. We used the IMB probe to separate the Brucella and labeled by the QD probe, and then detected the fluorescence intensity with a fluorescence spectrometer. The detection method takes 105 min with a limit of detection of 10 CFU/mL and ranges from 10 to 10 CFU/mL (R  = 0.9983), and it can be well used in real samples.

摘要

布鲁氏菌属是兼性胞内细菌,可在全球范围内引起人畜共患的布鲁氏菌病。传统的布鲁氏菌属检测方法需要48 - 72小时,不符合快速检测的需求。在此,基于多克隆抗体偶联量子点和抗体修饰磁珠开发了一种新的布鲁氏菌快速检测方法。首先,制备多克隆抗体IgG和IgY,然后分别将抗体与量子点(QDs)和免疫磁珠(IMB)偶联,它们通过N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺盐酸盐(EDC)和N-羟基琥珀酰亚胺(NHS)活化以形成探针。我们使用IMB探针分离布鲁氏菌并用QD探针标记,然后用荧光光谱仪检测荧光强度。该检测方法耗时105分钟,检测限为10 CFU/mL,线性范围为10至10 CFU/mL(R = 0.9983),并且可很好地用于实际样品检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fcf/5344867/f8a8846f5c51/11671_2017_1941_Fig1_HTML.jpg

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