| 猪繁殖与呼吸障碍综合征病毒(PRRSV)检测方法的建立 |
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| 引用本文： | 苏晓鸥,李玉荣,乔俊文,赵德明.猪繁殖与呼吸障碍综合征病毒(PRRSV)检测方法的建立[J].河北农业大学学报,2009,32(1). |
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| 作者姓名： | 苏晓鸥 李玉荣 乔俊文 赵德明 |
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| 作者单位： | 1. 中国农业大学动物医学院,国家动物海绵状脑病实验室,北京,100193
2. 河北农业大学,动物科技学院,河北,保定,071000 |
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| 基金项目： | 科技部资助项目，北京市科学技术委员会资助项目 |
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| 摘 要： | 根据GeneBank中已经发表的PRRSV毒株的ORF6基因高度保守序列,设计并合成了一对特异性引物,建立了用于检测PRRSV的RT-PCR诊断方法。从感染PRRSV-QD的Marc-145细胞中提取病毒RNA,然后经RT-PCR扩增,获得预期266 bp的目的片断,而猪伪狂犬病毒(PRV)、猪细小病毒(PPV)、猪圆环病毒(PCV)细胞培养物及Marc-145细胞进行同条件检测,结果为阴性;PRRSV扩增产物测序结果与GeneBank中已发表的PRRSV毒株序列同源性达94.4%~97.4%且应用本研究检测方法对已知基因型的10株PRRSV盲检,检出率100%。上述研究结果表明,所建立的RT-PCR诊断方法特异性好、敏感性高,可进一步应用于猪繁殖与呼吸综合征的临床诊断。
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| 关 键 词： | 猪繁殖与呼吸综合征 PRRSV ORF6 RT-PCR 检测 |
The establishment of detecting method for porcine reproductive and respiratory syndrome virus(PRRSV) |
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| SU Xiao-ou,LI Yu-rong,QIAO Jun-wen,ZHAO De-ming.The establishment of detecting method for porcine reproductive and respiratory syndrome virus(PRRSV)[J].Journal of Agricultural University of Hebei,2009,32(1). |
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| Authors: | SU Xiao-ou LI Yu-rong QIAO Jun-wen ZHAO De-ming |
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| Institution: | 1.National Animal Transmissible Spongiform Encephalopathy Laboratory;College of Veterinary Medicine;China Agricultural University;Beijing 100193;China;2.College of Animal Science and Technology;Agricultural University of Hebei;Baoding 071000;China |
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| Abstract: | Based on highly conservative ORF6 gene sequence of PRRSV on GeneBank,a pair of primers was designed and synthesized.The RT-PCR method for detecting PRRSV was established.PRRSV-QD RNA was extracted from virus-infected Marc-145 cell and then amplified by RT-PCR.The positive 266bp specific fragment was obtained as expected.Under the same conditions,however,the results for detection of PRV,PPV,PCV and Marc-145 cell by RT-PCR were all negative.The amplified sequence of PCR product of PRRSV-QD shares 94.4%-97.4% ... |
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| Keywords: | PRRSV ORF6 RT-PCR |
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