|
|||||
|
|
| 多重RT-PCR检测猪传染性胃肠炎病毒和猪流行性腹泻病毒 | |
| 引用本文： | 陈芳,刘惠莉,孙红立,曹祥荣,李震,赵立平.多重RT-PCR检测猪传染性胃肠炎病毒和猪流行性腹泻病毒[J].中国兽医学报,2005,25(1):9-12. |
| 作者姓名： | 陈芳 刘惠莉 孙红立 曹祥荣 李震 赵立平 |
| 作者单位： | 1. 上海市农业遗传育种重点实验室,上海,201106;南京师范大学,生命科学学院,江苏,南京,210097 2. 上海市农业科学院,畜牧兽医研究所,上海,201106;上海市农业遗传育种重点实验室,上海,201106 3. 南京师范大学,生命科学学院,江苏,南京,210097 4. 上海交通大学,生命科技学院,上海,200030 |
| 基金项目： | 上海市农委科技攻关项目 |
| 摘 要： | 为了检测、区分猪传染性胃肠炎(TGE)和猪流行性腹泻(PED)，建立了同时检测这2种疾病痛原体的多重PCR方法。参考GenBank登录的TGEV和PEDV纤突蛋白S基因序列，经DNAsis 2．0比较分析。分别筛选出TGEV、PEDVS基因相对保守区。以TGEV TH-98株(GenBank登陆号为AF494337)和PEDV CV777株(GenBank登陆号为NC003436)为参考模板，利用软件Primer3设计合成了2对寡核苷酸引物，以ST细胞培养的TGEV和PEDV毒株核酸为模板。利用所设计的2对引物进行多重RT-PCR扩增，结果同时得到与试验设计相符的499bp(TGEV)和199bp(PEDV)的扩增条带。而对其他3种猪病原的扩增结果均为阴性。敏感性试验结果表明。建立的多重RT-PCR方法可检测出10pgTGEV RNA和100pg PEDV RNA。 |
| 关 键 词： | TGEV 猪流行性腹泻病毒 猪传染性胃肠炎病毒 RT-PCR检测 引物 纤突蛋白 猪病 扩增 RNA 阴性 |
| 文章编号： | 1005-4545(2005)01-0009-04 |
Differentiation Between Porcine Epidemic Diarrhea Virus and Transmissible Gastroenteritis Virus by Multiplex RT-PCR |
|
| CHEN Fang.Differentiation Between Porcine Epidemic Diarrhea Virus and Transmissible Gastroenteritis Virus by Multiplex RT-PCR[J].Chinese Journal of Veterinary Science,2005,25(1):9-12. | |
| Authors: | CHEN Fang |
| Abstract: | A multiplex RT-PCR method was developed to simultaneously detect two pathogen of Porcine-TGEV and PEDV.According to conservative sequence of the spike protein gene of TGEV and PEDV at the GenBank,two sets of specific primers were designed by the software Primer 3.TGEV and PEDV RNA was extracted and amplified by the multiplex RT-PCR using these two sets of primers.The result showed that the two specific fragments of TGEV 499 bp and PEDV 199 bp were amplified,but negative results were gotten with other three porcine pathogenic viruses.The sensitivity assay indicated that 10 pg of TGEV and 100 pg PEDV RNA could be detected using this method. |
| Keywords: | TGEV PEDV multiplex RT-PCR detection |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |