鳜鱼病毒PCR诊断方法的建立
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中国水产科学研究院珠江水产研究所.广东广州510380

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农业部和广东省科委以及广东省自然科学基金资助( 980907)


Polymerase chain reaction ( PCR) amplification on diagnosis of Siniperca chuatsi virus disease
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Pearl River Fishery Research Institute, CAFS,Guangzhou 510380,China

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    摘要:

    从RAPD扩增的鳜鱼病毒(SCV)核酸电泳带中回收了二个片断,克隆子pUC19质粒(称为SCVE369和SCVE450),序列分析表明插入片段分别为369bp和450bp与GenBank序列没有显著的同源,根据克隆序列调计两对引物P1/P2和P3/P4 ,在健康鳜鱼,病鳜以及提纯的SCV核酸中进行PCR试验,结果表明,P1/P2组引物在SCV基因组中扩增出特异性核酸片段,可作为鳜鱼病毒PCR诊断,检测片段为369bp.

    Abstract:

    Two RAPD fragments of Siniperca chuatsi virus ( SCV) genome DNA recovered from agrose gel were inserted into plasmid pUC19 ( called SCVE369 and SCVE450) . The sequences reported the length of cloned f ragments were 369bp and 450bp. It doesn.. t show signif icant homology sequences against to those in GenBank. According those sequence two oligonucletide primersets ( P1/ P2 and P3/ P4) were designed and used to amplify genome DNA of SCV by the method of Polymerase Chain Reaction ( PCR) . The results showed primer ( P1/ P2) could specially amplify 369bp in size.. s fragment in SCV genome. Furthermore, this primer set was used to test the DNA samples including the normal mandarin f ish, cultured f ish naturally infected w ith SCV and the fish artif icially infected by SCV. The 369bp in size PCR product was only obtained in the mandarin fish carried SCV. Primer ( P1/ P2) can been used to diagnose SCV disease of mandarin f ish.

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李新辉.鳜鱼病毒PCR诊断方法的建立[J].水产学报,2001,25(1):

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  • 收稿日期:2014-04-16
  • 最后修改日期:2014-04-16
  • 录用日期:2014-04-16
  • 在线发布日期: 2014-04-16
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