套式RT2PCR方法检测临床样品中猪流感病毒.PDFVIP

套式 RTPCR 方法检测临床样品中的猪流感病毒 1 1 2 2 祁 　贤 ,陆承平 ,王贵平 ,宋长绪 ( 1. 南京农业大学 　农业部动物疫病诊断与免疫重点开放实验室 ,江苏南京 2 10095 ; 2 . 广东省农业科学院兽医研究所 ,广东广州 5 10640 ) Detection of Swine Influenza Virus in Clinical Samples Using RTnested PCR Assay Q I Xian1 ,L U Chengp ing1 ,WAN G Guiping2 ,SON G Changxun2 ( 1. Key laboratory of A nimal D isease D iag nostic and I m munology , M inist ry of ag ricult ure , N anj ing A g ricult ural University , N anj ing ) 210095 ; China; 2. Veterinary M edicine I nstit ute , Guang dong A g ricult ural A cademy of S cience , Guangz hou 510640 China Abstract : U sing t wo p air s of p rimer ba sed on con served sequence s of t he N P gene of influenza A vir u s , a R Tne st ed PCR a ssay wa s develop ed to det ect Swine influenza vir u s in clinical samp le s. No cro ss reaction s wit h t hree ot her swine p at ho gen s of re sp iratory di sea se such a s P orci ne resp i r atory an d rep rod uct i v e sy n d rom e v i r us , P orci ne ci rcov i r us t yp e 2 and M y cop l as m a s uip ne um oni ae . Wit h agaro se elect rop hore si s det ection , t he R Tne st ed PCR wa s 100 time s more sen sitive co mp ared to t hat of nonne st ed R TPCR . 35 clinical samp le s including 4 bloo d samp le s and 3 1 lung ti ssue s f ro m Sout her n China were t e st ed by R Tne st ed PCR , and 28 (80 %) out of 35 samp le were po sitive . By p rop agat ed in 9to 10dayol d SPF embryo nat ed chicken eggs , 23 st rain s of S w i ne i nf l uenz a v i r us were i solat ed f ro m 28 po sitive samp le s. The re sult of R Tne st ed