本试验旨在建立能同时检测猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)和猪A群轮状病毒(GAR)的多重RT-PCR检测方法。根据GenBank收录的PEDVM基因、TGEVN基因、GARVP7基因,利用软件Pri mer 5.0设计合成能分别特异性扩增PEDV、TGEV...本试验旨在建立能同时检测猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)和猪A群轮状病毒(GAR)的多重RT-PCR检测方法。根据GenBank收录的PEDVM基因、TGEVN基因、GARVP7基因,利用软件Pri mer 5.0设计合成能分别特异性扩增PEDV、TGEV、GAR相应基因的引物。利用这3对引物,作者建立了一种新的能够同时检测PEDV、TGEV和GAR的多重RT-PCR方法,并将这种方法和常规的RT-PCR进行了比较。实验室检测中,多重RT-PCR检测方法能够检测到35 pg的TGEV-PEDV-GAR三联苗的混合RNA。应用该方法检测了华中地区75份腹泻猪粪样,同时利用常规RT-PCR检测方法对该检测方法的敏感性和特异性进行了分析,结果表明该方法检测PEDV、TGEV、GAR的敏感性分别为92%、100%、100%,特异性均为100%。该方法敏感性高、特异性强,是一种新的检测PEDV、TGEV和GAR的方法。展开更多
DPO is one of the new specific primers for virus detection.In this study,five sets of dual priming oligonucleotide(DPO) primers for Potato virus X(PVX),Potato virus Y(PVY),Potato virus V(PVV),Tobacco ring spot virus(T...DPO is one of the new specific primers for virus detection.In this study,five sets of dual priming oligonucleotide(DPO) primers for Potato virus X(PVX),Potato virus Y(PVY),Potato virus V(PVV),Tobacco ring spot virus(TRSV) and Cucumber mosaic virus(CMV) were designed and applied to the detection of the main potato viruses with multiplex reverse transcription polymerase chain reaction(m-RT-PCR),including two quarantine viruses in China(PVV and TRSV).The results showed that DPO-m-RT-PCR exhi-bited high PCR specificity and sensitivity even under less than optimal PCR condition compared with conventional m-RT-PCR.DPO-m-RT-PCR could be applied in the healthy evaluation of seed potato and its exit-entry quarantine.展开更多
文摘本试验旨在建立能同时检测猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)和猪A群轮状病毒(GAR)的多重RT-PCR检测方法。根据GenBank收录的PEDVM基因、TGEVN基因、GARVP7基因,利用软件Pri mer 5.0设计合成能分别特异性扩增PEDV、TGEV、GAR相应基因的引物。利用这3对引物,作者建立了一种新的能够同时检测PEDV、TGEV和GAR的多重RT-PCR方法,并将这种方法和常规的RT-PCR进行了比较。实验室检测中,多重RT-PCR检测方法能够检测到35 pg的TGEV-PEDV-GAR三联苗的混合RNA。应用该方法检测了华中地区75份腹泻猪粪样,同时利用常规RT-PCR检测方法对该检测方法的敏感性和特异性进行了分析,结果表明该方法检测PEDV、TGEV、GAR的敏感性分别为92%、100%、100%,特异性均为100%。该方法敏感性高、特异性强,是一种新的检测PEDV、TGEV和GAR的方法。
文摘DPO is one of the new specific primers for virus detection.In this study,five sets of dual priming oligonucleotide(DPO) primers for Potato virus X(PVX),Potato virus Y(PVY),Potato virus V(PVV),Tobacco ring spot virus(TRSV) and Cucumber mosaic virus(CMV) were designed and applied to the detection of the main potato viruses with multiplex reverse transcription polymerase chain reaction(m-RT-PCR),including two quarantine viruses in China(PVV and TRSV).The results showed that DPO-m-RT-PCR exhi-bited high PCR specificity and sensitivity even under less than optimal PCR condition compared with conventional m-RT-PCR.DPO-m-RT-PCR could be applied in the healthy evaluation of seed potato and its exit-entry quarantine.
