PCV2和PCV3双重纳米PCR方法的建立及初步应用

发布时间：2019-01-12 06:55

【摘要】：本研究旨在建立一种同时检测猪圆环病毒2型(PCV2)和猪圆环病毒3型(PCV3)的双重纳米PCR(nanodPCR)方法,同时应用该方法对PCV2和PCV3进行检测。参考GenBank中登录的PCV2和PCV3型基因序列分别设计特异性引物,对nano-dPCR的反应条件进行优化;同时考察所建立的nano-dPCR检测方法的特异性和敏感性。结果显示,所建方法的特异性和敏感性良好,对PCV2和PCV3两种病毒的最低核酸检测量分别为93.2和91.6拷贝/μL,其敏感性比普通PCR高100倍。应用该方法对送检的265份临床样本进行检测,结果显示,PCV2和PCV3在猪群中存在普遍感染,PCV3和PCV2的阳性率分别为16.6%和14.7%,混合感染阳性率为6.8%。临床样品的检测结果表明,本试验建立的nano-dPCR方法为PCV2和PCV3早期感染进行快速、敏感鉴别诊断提供了新方法。
[Abstract]:The purpose of this study was to establish a double nano PCR (nanodPCR) method for simultaneous detection of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3), and to detect PCV2 and PCV3 simultaneously. Specific primers were designed to optimize the reaction conditions of nano-dPCR with reference to the PCV2 and PCV3 gene sequences recorded in GenBank, and the specificity and sensitivity of the established nano-dPCR detection method were investigated. The results showed that the specificity and sensitivity of the proposed method were 93.2 and 91.6 copies / 渭 L for PCV2 and PCV3, respectively. The sensitivity of the method was 100 times higher than that of normal PCR. This method was used to detect 265 clinical samples. The results showed that PCV2 and PCV3 were commonly infected in pigs. The positive rates of PCV3 and PCV2 were 16.6% and 14.70.The positive rate of mixed infection was 6.8%. The detection results of clinical samples show that the nano-dPCR method established in this study provides a new method for the rapid and sensitive differential diagnosis of early infection of PCV2 and PCV3.
【作者单位】：中国农业科学院北京畜牧兽医研究所;农业部兽用药物与诊断技术北京科学观测实验站;青岛机场出入境检验检疫局;
【基金】：中国农业科学院基本科研业务费(2016ywf-yb-12);中国农业科学院创新工程(ASTIP-IAS15)
【分类号】：S852.651

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