塞内卡病毒A的RT-PCR检测方法的建立和应用

doi:10.11843/j.issn.0366-6964.2019.06.022

摘要/Abstract

摘要： 塞内卡病毒A（Senecavirus A，SVA）是一种无囊膜的单股正链RNA病毒，可引起成年母猪水疱性病变，并导致新生仔猪死亡。为建立SVA的快速检测方法，本研究对NCBI发表的40株SVA的基因序列进行同源比较，选择保守区域设计了3对引物P1、P2和P3，从中筛选出最佳扩增引物，通过对引物浓度、退火温度、延长时间、循环次数进行优化，成功建立了SVA的RT-PCR检测方法。用该方法检测SVA、脑心肌炎病毒（EMCV）、口蹄疫病毒（FMDV）、猪伪狂犬病病毒（PRV）、猪繁殖与呼吸综合征病毒（PRRSV）、猪瘟病毒（CSFV）和猪流行性腹泻病毒（PEDV）无交叉反应，具有良好的特异性。敏感性试验结果显示，病毒检测限可达1 TCID₅₀，比国外文献报道的检测方法灵敏性更高。用该方法检测山东省的100份猪组织样品，阳性率为2%。本研究所建立的方法特异性强、敏感性高、可靠性好，为SVA的快速检测及流行病学调查提供了有效的技术手段。

Abstract: Senecavirus A(SVA) is a nonenveloped, single-stranded RNA virus, which can cause vesicular lesions in sows and newborn piglet death. In order to establish a rapid assay for detection of SVA, we compared the genetic sequences of 40 SVA genes published on NCBI, and the best primers was selected from three pairs of primers which designed based on the conserved regions of sequences. The RT-PCR detection method of SVA was established successfully after the optimization of primer concentration, annealing temperature, extension time and cycle number. SVA, Encephalomyocarditis virus (EMCV), foot-and-mouth disease virus (FMDV), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), swine fever virus (CSFV), porcine epidemic diarrhea virus (PEDV) were detected by RT-PCR, which showed a good specificity with no cross-reaction. The sensitivity test was carried out as well, and sensitivity of the detection could reach up to 1 TCID₅₀, which shows it is more sensitive than the methods reported in foreign literatures. One hundred samples of pig tissue were detected using this method, and the positive rate was 2%. The establishment of this method, with its great specificity, high sensitivity and ideal reliability, could provide an effective technical means for SVA detection and epidemiological investigation.

中图分类号:

S852.659.6

范慧, 李亮, 姜平, 王先炜, 李玉峰, 白娟. 塞内卡病毒A的RT-PCR检测方法的建立和应用[J]. 畜牧兽医学报, 2019, 50(6): 1312-1318.

FAN Hui, LI Liang, JIANG Ping, WANG Xianwei, LI Yufeng, BAI Juan. Establishment and Application of a RT-PCR Assay for Detection of Senecavirus A(SVA)[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1312-1318.

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