[1]张鹏超,师小潇,哈 卓,等.猪流感病毒SYBR GreenⅠ定量RT-PCR检测方法的建立及应用[J].中国预防兽医学报,2010,(10):768-722.[doi:10.3969/j.issn.1008-0589.2010.10.06]
 ZHANG Peng-chao,SHI Xiao-xiao,HA Zhuo,et al.Establishment of SYBR GreenⅠreal time RT-PCR for detecting swine influenza virus[J].Chinese journal of preventive veterinary medicine,2010,(10):768-722.[doi:10.3969/j.issn.1008-0589.2010.10.06]
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猪流感病毒SYBR GreenⅠ定量RT-PCR检测方法的建立及应用
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2010年10期
页码:
768-722
栏目:
诊断和检测技术
出版日期:
2010-10-01

文章信息/Info

Title:
Establishment of SYBR GreenⅠreal time RT-PCR for detecting swine influenza virus
文章编号:
1008-0589(2010)10-0768-05
作者:
张鹏超12师小潇13哈 卓13刘永杰2刘惠莉13*
1. 上海市农业科学院 畜牧兽医研究所,上海 201106;
2. 南京农业大学 动物医学院,江苏 南京 210095;
3. 上海市农业遗传育种重点实验室,上海 201106
Author(s):
ZHANG Peng-chao12 SHI Xiao-xiao13 HA Zhuo13 LIU Yong-jie2 LIU Hui-li13*
1. Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai Municipal Key Laboratory of Agricultural Genetics and Breeding, Shanghai 201106, China;
2. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;
3. Shanghai Muncicpal Key Laboratory of Agricultural Genetics and Breeding, Shanghai 201106, China
关键词:
猪流感病毒SYBR Green I荧光定量PCR方法组织分布
Keywords:
swine influenza virus real-time fluorescent quantitative PCR virus distribution
分类号:
S852.65
DOI:
10.3969/j.issn.1008-0589.2010.10.06
文献标志码:
A
摘要:
为研究猪流感病毒(SIV)感染后SIV在体内的分布规律,本研究设计针对SIV NP基因保守区引物,建立了SIV SYBR Green I荧光定量PCR方法。该方法对SIV核酸检测灵敏度为30 TCID50,对健康猪肺组织cDNA、猪瘟病毒(CSFV)、猪蓝耳病病毒(PRRSV)核酸呈阴性反应。采用建立的荧光定量RT-PCR方法对人工感染A/swine/Shanghai/1/2007/H1N2 (Sw/SH/1/2007)猪进行检测,结果显示:病毒在猪鼻腔粘膜持续存在至感染后第8 d;肛门拭子在感染后2 d~8 d可持续检测到病毒核酸;喉头、气管在感染后第3 d可检到病毒核酸,并持续至第10 d;肺部淋巴结、脾脏在感染后5 d~7 d检测到病毒核酸阳性;其它脏器均未检测到病毒核酸,从而确定呼吸道系统及脾脏是SIV定殖的主要场所,SIV感染猪后向外排毒周期约为1周。
Abstract:
The distribution of swine influenza virus (SIV) in pigs was investigated after artificially infected 40-day-old pigs with A/Sw/SH/1/2007 SIV strain. The infected pigs showed clinical symptom of cough and sneezing, and recovered within two weeks. The nasal swabs, anal swabs and tissue samples were taken at selected time points post inoculation (DPI) and detected for SIV by fluorescent quantitative PCR. Nucleic acid of the virus could be detected in nasal swabs immediately after inoculation and persistent for 8 days; The virus was detected positive in respiratory mucus from 3 DPI to 10 DPI, in the anal swabs from 2 DPI to 8 DPI, and in the lung lymph notes and spleen was 5 DPI to 7 DPI. It was concluded that the A/Sw/SH/1/2007 virus was mainly distributed in respiratory system and spleen, and the respiratory mucous is the main port of swine influenza virus colonization.

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备注/Memo

备注/Memo:
收稿日期:2009-12-13
基金项目:上海市农委重点攻关项目(沪农科攻字(2004)第12-2号、沪农科攻字(2009)第5-2号)
作者简介:张鹏超(1985-),男,山东莱芜人,硕士研究生,主要从事动物病毒学研究.
*通信作者:E-mail:huilil@163.com
更新日期/Last Update: 2010-09-27