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用于快速现场检测非洲猪瘟病毒的基于量子点的免疫层析试纸条的研制。

Development of a quantum dots based immunochromatographic strip for rapid and on-site detection of African swine fever virus.

作者信息

Wu Yuxiang, Wang Changjiang, Yu Jinzhi, Ma Fengyun, Liu Jie, Tan Jing, Qu Guanggang

机构信息

School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo, China.

Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou, China.

出版信息

Microb Pathog. 2024 Jun;191:106669. doi: 10.1016/j.micpath.2024.106669. Epub 2024 May 1.

Abstract

African swine fever (ASF) is a lethal disease caused by ASF virus (ASFV), severely impacting the global swine industry. Though nuclear acid-based detection methods are reliable, they are laboratory-dependent. In this study, we developed a device-independent, user friendly and cost-effective quantum dots based immunochromatographic strip (QDs-ICS) with high specificity and sensitivity for the rapid and on-site detection of ASFV antigen. For the preparation of the QDs-ICS, we generated a monoclonal antibody (mAb) mAb-8G8 and polyclonal antibody (pAb) against ASFV-p72 protein. The pAb was labelled with QDs to be used as the detection probe and the mAb-8G8 was coated on the nitrocellulose membrane as the test line. Our results proved that the strip displayed no cross-reactivity with other swine viruses and detection limit of the QDs-ICS was down to 1 ng/mL for the ASFV-p72 protein with great reproducibility. The strip also exhibited high stability with a storage period up to 12 months under room temperature. Twenty blind samples and one hundred clinical samples were examined by the QDs-ICS, conventional PCR and real-time PCR method, respectively. Results showed that the agreement rate between the QDs-ICS and PCR method was 100%, and the agreement rate between the strip and real-time PCR was 94%. The novel QDs-ICS developed here would be an effective tool for on-site detection of ASFV.

摘要

非洲猪瘟(ASF)是一种由非洲猪瘟病毒(ASFV)引起的致命疾病,对全球养猪业造成严重影响。尽管基于核酸的检测方法可靠,但它们依赖于实验室。在本研究中,我们开发了一种独立于设备、用户友好且经济高效的基于量子点的免疫层析试纸条(QDs-ICS),用于快速现场检测ASFV抗原,具有高特异性和灵敏度。为了制备QDs-ICS,我们制备了针对ASFV-p72蛋白的单克隆抗体(mAb)mAb-8G8和多克隆抗体(pAb)。将pAb用量子点标记作为检测探针,将mAb-8G8包被在硝酸纤维素膜上作为检测线。我们的结果证明,该试纸条与其他猪病毒无交叉反应,对于ASFV-p72蛋白,QDs-ICS的检测限低至1 ng/mL,具有良好的重复性。该试纸条在室温下储存期长达12个月时也表现出高稳定性。分别用QDs-ICS、常规PCR和实时PCR方法检测了20份盲样和100份临床样品。结果表明,QDs-ICS与PCR方法的符合率为100%,试纸条与实时PCR的符合率为94%。这里开发的新型QDs-ICS将是现场检测ASFV的有效工具。

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