[1]樊晓旭?,宋翥远?,赵永刚,等.塞尼卡谷病毒重组酶聚合酶扩增-侧流层析试纸条检测方法的建立[J].中国预防兽医学报,2018,(05):406-410.[doi:0.3969/j.issn.1008-0589.201708027]
 FAN Xiao-xu,SONG Zhu-yuan,ZHAO Yong-gang,et al.Establishment of recombinase polymerase amplification-lateral flow dipstick for the detection of Seneca valley virus[J].Chinese journal of preventive veterinary medicine,2018,(05):406-410.[doi:0.3969/j.issn.1008-0589.201708027]
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塞尼卡谷病毒重组酶聚合酶扩增-侧流层析试纸条检测方法的建立()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年05
页码:
406-410
栏目:
诊断和检测技术
出版日期:
2018-05-15

文章信息/Info

Title:
Establishment of recombinase polymerase amplification-lateral flow dipstick for the detection of Seneca valley virus
文章编号:
1008-0589(2018)5-0406-05
作者:
 

樊晓旭1?宋翥远2?赵永刚1赵 明3董雅琴4张永强1李园丽15迟田1刘春菊1戈胜强1张志诚1吴晓东1王树双1王志亮1*

 (1. 中国动物卫生与流行病学中心 外来病研究中心,山东 青岛266032;2. 青岛市动物园管理处,山东 青岛266071;
3. 青岛市动物疫病预防控制中心,山东 青岛 266100;4. 中国动物卫生与流行病学中心 畜病监测室,山东 青岛 266032;
5. 沈阳农业大学 畜牧兽医学院,辽宁 沈阳 110866)
Author(s):
 

FAN Xiao-xu1? SONG Zhu-yuan2? ZHAO Yong-gang1 ZHAO Ming3 DONG Ya-qin4 ZHANG Yong-qiang1 LI Yuan-li15 CHI Tian-ying1 LIU Chun-ju1 GE Sheng-qiang1 ZHANG Zhi-cheng1 WU Xiao-dong1 WANG Shu-shuang1 WANG Zhi-liang1*

 (1. National Research Center for Exotic Animal Disease, China Animal Health and Epidemiology Center, Qingdao 266032, China;
2. Management Office of Qingdao Zoo, Qingdao 266071, China; 3. Qingdao Animal Disease Control Center, Qingdao 266100, China;
4. Division of livestock diseases surveillance, China Animal Health and Epidemiology Center, Qingdao 266032, China;
5. College of animal husbandry and veterinary medicine, Shenyang Agricultural University, Shenyang 110866, China
关键词:
塞尼卡谷病毒重组酶聚合酶扩增侧向流试纸条
Keywords:
 seneca valley virus  recombinase polymerase amplification  lateral flow dipstick
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201708027
文献标志码:
A
摘要:
为建立塞尼卡谷病毒(SVV)重组酶聚合酶扩增-侧流层析试纸条(RPA-LFD)检测方法,本研究针对SVV 3D基因设计了引物和探针,通过反应条件优化、特异性、灵敏性和重复性试验,建立了SVV RPA-LFD检测方法。特异性试验结果显示,该方法与口蹄疫病毒、猪瘟病毒、猪繁殖与呼吸道综合征病毒、猪流行性腹泻病毒、猪传染性胃肠炎病毒、圆环病毒、伪狂犬病毒无交叉反应;在40 ℃、20 min内可检测的最低浓度为56拷贝/μL质粒标准品;通过3次重复试验检测,LFD检测线灰度值变异系数范围在4.49 %~9.73 %。利用该方法对120份国内临床样品进行检测,结果均为阴性。本研究首次建立了检测SVV的等温、快速RPA-LFD方法,读取结果不依赖任何仪器设备,为猪群感染SVV的现场快速诊断、流行病学研究和根除方案的制定提供帮助。
  
Abstract:
Recombinase polymerase amplification (RPA) is an isothermal and rapid amplification technique for the detection of nucleic acid developed recently. Along with lateral flow dipstick (LFD), the result of the amplicon can be read-out directly. In this study, a RPA-LFD combined protocol was established with the primers and probes targeting seneca valley virus (SVV) 3D gene for SVV rapid detection. Under the optimized detection conditions, the assay was able to complete the detection within 20 min at 40 ℃, which was specificity for SVV detection and had no cross reaction with the foot and mouth disease virus, classicalswine fever virus, porcine reproductive and respiratory syndrome virus, porcine epidemic diarrhea virus, transmissible gastroenteritis virus, porcine circovirus and pseudorabies virus. The sensitive of this assay was capable to detect out of 56 copies/μL of SVV. The coefficient of variation regarding the gray values of test lines was in the range of 4.49% to 9.73% assayed from seven series diluted samples (5.6×107 to 5.6×101 copies/μL) with three repeat detections. A total of 120 samples were detected by this method, and the results were all negative. The established isothermal rapid amplification assay for the detection of SVV facilitates the on-site rapid diagnosis, epidemiological study and eradication of SVV among pigs.
更新日期/Last Update: 2018-05-15