基于纳米磁珠和量子点的核酸传感器检测猪瘟病毒  

Detection of Classical Swine Fever Virus by Nucleic Acid Sensor Based on Magnetic Beads and Quantum Dots

作　　者：赵妍 吴梦凡 雷蕾 潘兹书[1] ZHAO Yan;WU Mengfan;LEI Lei;PAN Zishu(College of Life Sciences, Wuhan University, Wuhan 430072, Hubei,China)

机构地区：[1]武汉大学生命科学学院

出　　处：《武汉大学学报（理学版）》2019年第4期411-416,共6页Journal of Wuhan University:Natural Science Edition

基　　金：国家重点研发计划(2016YFD0500708)

摘　　要：通过比对猪瘟病毒(classical swine fever virus,CSFV)基因组核苷酸序列,筛选出CSFV的保守靶序列。设计针对该靶序列的分子探针,通过生物素与链霉亲和素的相互作用,将捕获探针偶联到纳米磁珠,将检测探针偶联到荧光量子点,建立了基于纳米磁珠分离富集靶标和量子点探针捕获特性的生物传感器检测CSFV核酸的方法。该方法对标准品的检出限为104copies/μL,对临床样本的检出限为106copies/μL,且与其他猪病病毒无交叉反应,具有良好的特异性。该生物传感器对25份临床样本的检测结果与荧光定量PCR方法检测结果相符。The target viral sequences were determined based on alignment of genome sequences of classical swine fever virus(CSFV) and the molecluar probes were designed direct to the target sequences. The quantum dots(QD) conjugated with detection probes and the magnetic beads(MB) conjugated with capture probes were prepared by interaction between biotin and streptavidin, respectively. The target viral RNAs were separated and enriched from nanomagnetic beads and captured by quantum dot fluorescent signals. A biosensor for detecting CSFV was developed. As low as 10~4 copies/μL viral RNA could be detected and no cross-reactivity could be observed for other swine viruses. The detection limit of CSFV for clinical specimens was 10~6 copies/μL, exhibiting a high degree of specificity. The results of biosensor detection were in consistent with the quantitative PCR assay for 25 clinical specimens.

关 键 词：猪瘟病毒 量子点 纳米磁珠 核酸传感器 

分 类 号：Q93[生物学—微生物学]