[1]冯瑜菲,赵国辉,徐青元,等. 蓝舌病病毒一步RT-PCR检测方法的建立[J].中国预防兽医学报,2014,(09):712-714.[doi:10.3969/j.issn.1008-0589.2014.09.12]
 FENG Yu-fei,ZHAO Guo-hui,XU Qing-yuan,et al.Development of one-step RT-PCR method for the detection of bluetongue virus[J].Chinese journal of preventive veterinary medicine,2014,(09):712-714.[doi:10.3969/j.issn.1008-0589.2014.09.12]
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 蓝舌病病毒一步RT-PCR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:
2014年09期
页码:
712-714
栏目:
诊断和检测技术
出版日期:
2014-09-15

文章信息/Info

Title:
Development of one-step RT-PCR method for the detection of bluetongue virus
文章编号:
1008-0589(2014)09-0712-03
作者:
 冯瑜菲12赵国辉2徐青元2杨 涛2孙恩成2李俊平2吴东来12*
 1. 东北农业大学 动物医学学院,黑龙江 哈尔滨150030;2. 中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室/
农业部兽医公共卫生重点开放实验室,黑龙江 哈尔滨 150001
Author(s):
 FENG Yu-fei12 ZHAO Guo-hui2 XU Qing-yuan2 YANG Tao2 SUN En-cheng2LI Jun-ping2 WU Dong-lai12*
 1. College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China; 2. Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China
关键词:
 蓝舌病病毒一步RT-PCR分子诊断
Keywords:
 bluetongue virus (BTV)  one-step RT-PCR  molecular diagnostics
分类号:
S852.65
DOI:
10.3969/j.issn.1008-0589.2014.09.12
文献标志码:
A
摘要:
 为建立蓝舌病病毒(BTV)群特异性核酸检测方法,本研究针对BTV S10基因保守区域设计并合成1对特异性引物,建立了一步RT-PCR检测方法,并分别进行了特异性试验、敏感性试验以及临床样品的检测。实验结果表明:该方法对BTV1~24型均有特异性反应,而对茨城病毒(IBAV)、中山病毒(CV)和赤羽病病毒(AKAV)无交叉反应,具有良好的特异性;最低检出量为102 TCID50/mL。 此外,该方法能有效的从羊的抗凝血样品中检测出BTV 核酸。本研究所建立的一步RT-PCR检测方法可以用于BTV的快速检测及流行病学调查。
Abstract:
 To establish the assay for detecting bluetongue virus (BTV), one-step RT-PCR assay was developed with a pair of group specific primers targeting the sequence of BTV S10 gene for the detection of BTV from serotype 1 to 24. The test results showed that the one-step RT-PCR was specific for detection the serotype 1 to 24 of BTV with a detection limit of 102 TCID50/mL, but no cross amplification for IBAV, CV and AKAV. In addition, this assay was capable to detect BTV from the clinical blood sample of sheep. These results demonstrated the established one-step RT-PCR had the potential for the rapid detection and epidemic surveys of BTV.

参考文献/References:

[1] Al Ahmad M Z, Bruyas J F, Pellerin J L, et al. Evaluation of bluetongue virus (BTV) decontamination techniques for caprine embryos produced in vivo [J]. Theriogenology, 2012, 78: 1286- 1293.
[2]Maan N S, Maan S, Belaganahalli M N, et al. Identification and differentiation of the twenty six bluetongue virus serotypes by RT-PCR amplification of the serotype-specific genome segment 2[J]. PLoS One, 2012, 7: e32601.
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[5]Wei Peng, Sun En-cheng, Liu Ni-hong, et al. Identification of a novel bluetongue virus 1-specific B-cell epitope using a monoclonal antibody against the VP2 protein[J]. Arch Virol, 2013, 158: 1099-1104.

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备注/Memo

备注/Memo:
收稿日期:2014-03-11
基金项目:中央级公益性科研院所基本科研业务费专项(030201310)
作者简介:冯瑜菲(1984-),女,黑龙江哈尔滨人,博士研究生,主要从事分子病毒学研究.
*通信作者:E-mail:dlwu@hvri.ac.cn
更新日期/Last Update: 2014-09-18