[1]崔尚金,胡泉博,刘业兵*.非洲猪瘟病毒高效纳米PCR检测方法的建立及初步应用[J].中国预防兽医学报,2012,(10):807-809.[doi:doi: 10.3969/j.issn.1008-0589.2012.10.12]
 CUI Shang-jin,HU Quan-bo,LIU Ye-bing*.The development of nanometer PCR assay for detectionof the Africa swine fever virus[J].Chinese journal of preventive veterinary medicine,2012,(10):807-809.[doi:doi: 10.3969/j.issn.1008-0589.2012.10.12]
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非洲猪瘟病毒高效纳米PCR检测方法的建立及初步应用
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2012年10期
页码:
807-809
栏目:
诊断和检测技术
出版日期:
2012-10-15

文章信息/Info

Title:
The development of nanometer PCR assay for detection
of the Africa swine fever virus
文章编号:
1008-0589(2012)10-0807-03
作者:
崔尚金1胡泉博1刘业兵2*
(1. 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/猪传染病研究室, 黑龙江 哈尔滨 150001;
2. 中国兽医药品监察所,北京 100081)
Author(s):
CUI Shang-jin1 HU Quan-bo2 LIU Ye-bing2*
(1. Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute,
Chinese Academy of Agricultural Sciences, Harbin 150001, China; 2. China Institute of Veterinary Drug Control, Beijing 100081, China)
关键词:
纳米PCR非洲猪瘟病毒P54基因
Keywords:
nanometer-PCR Africa swine fever virus P54 gene
分类号:
S852.65
DOI:
doi: 10.3969/j.issn.1008-0589.2012.10.12
文献标志码:
A
摘要:
为将纳米PCR技术引入非洲猪瘟病毒(ASFV)检测,本研究建立了ASFV的高效纳米PCR检测方法,该方法采用纳米金颗粒作为热导介子,提高了PCR反应效率,采用该方法同时检测ASFV、大肠杆菌、猪伪狂犬病毒、猪圆环病毒2型、典型猪瘟病毒、猪繁殖与呼吸综合征病毒、猪捷申病毒、猪脑心肌炎病毒等,结果只有ASFV能够扩增出552 bp的目的片段。敏感性试验表明纳米PCR检测技术是常规PCR检测技术敏感性的1 000倍以上,最低核酸拷贝数检出量可以达到10 个拷贝。采用该方法对黑龙江、吉林和河南3个省份的8个地区临床送检的69份样品进行检测,检测结果显示均为阴性,在以上3个地区均未发现ASFV感染情况。
Abstract:
In the present study, a nanometer PCR (nano-PCR) assay was developed to detect the Africa swine fever virus (ASFV) using the P54 gene containing recombinant plasmid as the tamplate for 552bp fragment amplification. The amplification was efficiently enhanced with the gold nanoparticles as thermal mediator in the amplification system, and the sensitivity of the nano-PCR was more than 1,000 times than that of conventional PCR with a detection limit of 10 copies and had no cross-reactions with E. coli, porcine pseudorabies virus, porcine circovirus type II, classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine Teschovirus 8 and encephalomyocarditis virus. Furthermore, a total of 69 clinical samples from Heilongjiang, Jilin and Henan provinces were detected by this assay. The results showed that all the samples were negative, indicating no ASFV infection occurred in these areas.

参考文献/References:

[1]曲连东,于康震. 非洲猪瘟研究进展[J]. 中国兽医科技,1998,28 (11):42-43.
[2]王华,王君玮,徐天刚,等. 非洲猪瘟流行病学和诊断方法的研究进展[J]. 中国兽医科学,2008,38(6):544-548.
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[4]Mckillen J, Hjertner B, Millar A, et al. Molecular beacon real- time PCR detection of swine viruses [J]. J Virol Methods, 2007, 140(1-2): 155-165.
[5]Li Hui-xiang, Rothberg L. Colorimetric detection of DNA sequ- ences based on electrostatic interactions with unmodified gold nanoparticles [J]. PNAS, 2004, 101(39): 14036-14039.
[6]侯艳梅,董志珍,赵祥平,等. 非洲猪瘟病毒P54基因原核表达载体的构建与表达[J]. 中国兽医科学,2009,39(04):316-320.
[7]李海阔,曹雪雁,周知磊,等. 纳米金增强复杂体系中PCR扩增低拷贝基因的反应特异性初步研究[J]. 中国生物工程杂志,2009,29 (8):75-80.
[8]米丽娟,朱红平,张晓东,等. “纳米粒子PCR”中纳米金与聚合酶相互作用机制探讨[J]. 科学通报,2007,52(8):893-897.
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备注/Memo

备注/Memo:
收稿日期:2012-05-12
基金项目:科技成果转化项目(2011GB2C600011)
作者简介:崔尚金(1971-),山东青州人,研究员,主要从事猪病病毒学诊断、流行病学与免疫学研究.
*通信作者:E-mail:liuyebing@ivdc.gov.cn
更新日期/Last Update: 2012-10-18