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| 猪瘟、猪圆环病毒病和猪繁殖与呼吸综合征寡核苷酸芯片诊断方法的建立与初步应用 | |
| 引用本文： | 叶芬,蔡家利,王昱,李应国.猪瘟、猪圆环病毒病和猪繁殖与呼吸综合征寡核苷酸芯片诊断方法的建立与初步应用[J].中国兽医学报,2011,31(10). |
| 作者姓名： | 叶芬 蔡家利 王昱 李应国 |
| 作者单位： | 1. 西南大学动物科技学院,重庆,400715 2. 重庆理工大学药学与生物工程学院,重庆,400054 3. 重庆市出入境检验检疫局,重庆,400020 |
| 基金项目： | 重庆市科技攻关资助项目(2009CC05) |
| 摘 要： | 猪繁殖与呼吸综合征病毒（PRRSV）、猪瘟病毒（CSFV）和猪圆环病毒Ⅱ型（PCV-2）是引发猪繁殖障碍的主要病原,建立其基因芯片快速检测体系,对开发猪繁殖障碍快速检测试剂盒具有重要意义。根据GenBank中已发表的PRRSV、CSFV和PCV-2的病毒基因组序列,设计合成特异性引物和特异性较强的60mer的寡核苷酸探针,并将探针按所设计阵列固定于表面经氨基化修饰的玻片上,制备出寡核苷酸芯片。通过引物标记及特异性验证,建立了带标记引物的多重PCR检测体系,从而产生大量可与寡核苷酸探针特异性互补的带标记的DNA片段。将标有荧光染料的扩增产物与芯片上寡核苷酸探针杂交,扫描、分析芯片上荧光信号。结果表明,芯片上各样本对应探针位点呈现阳性荧光信号,而阴性对照和空白对照则基本不能检测到荧光信号。分别用基因芯片检测方法和PCR/RT-PCR检测60份临床病料,两者符合率高达92%,表明该检测技术能够用于临床病料的检测及快速诊断PRRSV、CSFV和PCV-2。 |
| 关 键 词： | 多重PCR 基因芯片 PCV-2 CSFV PRRSV |
Establishment and application of oligonucleotide microarray diagnostic technique for detection of porcine reproductive and respiratory syndrome virus,classical swine fever virus and porcine circovirus type 2 virus |
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| YE Fen,CAI Jia-li,WANG Yu,LI Ying-guo.Establishment and application of oligonucleotide microarray diagnostic technique for detection of porcine reproductive and respiratory syndrome virus,classical swine fever virus and porcine circovirus type 2 virus[J].Chinese Journal of Veterinary Science,2011,31(10). | |
| Authors: | YE Fen CAI Jia-li WANG Yu LI Ying-guo |
| Institution: | YE Fen1,CAI Jia-li2*,WANG Yu3,LI Ying-guo3(1.College of Animal Science and Technology,Southwest University,Chongqing 400715,China,2.College of Pharmaceutical and Biological Engineering,Chongqing University of Technology,Chongqing 400054,3.Congqing Entry-Exit Inspection and Quarantine Bureau,Chongqing 400020,China) |
| Abstract: | Porcine reproductive and respiratory syndrome virus(PRRSV),classical swine fever virus(CSFV) and porcine circovirus type 2(PCV-2) are main pathogens of porcine reproduction barrier diseases.The establishment of the oligonucleotide microarray diagnostic technique is very important for developing porcine reproductive problems detection kit.Based on the similarity of the viral sequences deposited in the GenBank database,species-specific 60-mer oligonucleotide probes(oligoprobes) as the capture probes were desi... |
| Keywords: | multiplex PCR oligonucleotide microarray diagnostic technique porcine circovirus type 2 virus classical swine fever porcine reproductive and respiratory syndrome virus |
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