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

机译:一种带有量子点和磁珠结合不同多克隆抗体的布鲁氏菌快速检测方法

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摘要

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 103 CFU/mL and ranges from 10 to 105 CFU/mL (R 2 = 0.9983), and it can be well used in real samples.
机译:布鲁氏菌属是在世界范围内引起人畜共患布鲁氏菌病的兼性细胞内细菌。检测布鲁氏菌属的传统方法。花费48–72小时不满足快速检测的需要。在此,基于多克隆抗体缀合量子点和抗体修饰的磁珠,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,然后分别与量子点(QD)和免疫磁珠(IMB)缀合的抗体,这些量子点被N-(3-二甲基氨基丙基)-N'-乙基-cardibodiimide盐酸盐(EDC)激活)和N-羟基琥珀酰亚胺(NHS)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。检测方法需要105分钟,检出限为10 3 CFU / mL,范围为10到10 5 CFU / mL(R 2 = 0.9983),并且可以很好地用于实际样本中。

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