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《Acta Agriculturae Zhejiangensis》 2012-03
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Development of one-step real-time fluorescent quantitative RT-PCR assay for detection of porcine reproductory syndrome virus strains

LU Bin1,2,YUAN Xiu-fang1,XU Li-hua1,LI Jun-xing1,GUO Yong1,WANG Yi-cheng1,*(1Institute of Animal Husbandry and Veterinary,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;2 Animal Husbandry and Veterinary Bureau of Haiyan County of Zhejiang,Jiaxing 314300,China)  
To identify porcine reproductive and syndrome virus(PRRSV) rapidly using real-time RT-PCR technique,primers and TaqMan probe were designed based on the PRRSV sequence.The fluorescent quantitative RT-PCR assay for detection of PRRSV was established.Its specificity,sensitivity and stability were examined.The assay was also proven to be specific based on the results that no amplification was found by detecting RNA/DNA from PCV,JEV,SFV,PRV and PPV.10 copies of template RNA or one TCID50 of PRRSV could be detected.The coefficient variation(CV%) of intra/inter-assay for same DNA sample was less than 1%.72 samples were examined by the fluorescent quantitative RT-PCR and the conventional RT-PCR respectively.40 were detected to be infected with PRRSV by the real-time PCR,but only 28 were confirmed to be infected with PRRSV by conventional PCR.These results showed that the fluorescent quantitative RT-PCR assay was rapid,specific,sensitive and simple for the detection of PRRSV.
【Fund】: 浙江省重大科技专项农业项目(2008C12049)
【CateGory Index】: S858.28
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【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 Huang Changchun;Quanzhou Animal Disease Prevention and Control Center;;The epidemiological investigation of porcine reproductive and respiratory syndrome in Quanzhou city[J];福建畜牧兽医;2015-06
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【Co-references】
Chinese Journal Full-text Database 10 Hits
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