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Position: Home > Articles > Development and application of high sensitivity RT-PCR detection of arabis mosaic virus Acta Agriculturae Shanghai 2013,29 (5) 80-84

南芥菜花叶病毒高灵敏度检测方法的建立及应用

作者：

吴东妮;陈舜胜;杨翠云;于翠

单位：

上海出入境检验检疫局;上海海洋大学

关键词：

南芥菜花叶病毒;检测;RT-PCR;聚乙二醇;免疫磁珠;敏感度

摘要：

以感染ArMV的百合种球制备粗提液,分别用聚乙二醇(PEG-6000)和免疫磁珠(Immuno-magnetic beads,IMB)进行富集处理,以富集处理后的样品进行ELISA、RT-PCR和SYBR Green I实时荧光RT-PCR检测。结果表明:富集处理能显著提高这些方法从百合种球样品中检测ArMV的灵敏度。PEG-RT-PCR可检测稀释至10~(-5)的样品,IMB-RT-PCR可检测稀释至10~(-3)的样品。实时荧光RT-PCR检测也表明,PEG对百合种球中ArMV的富集效果优于IMB处理。对19批进境百合种球进行ArMV的RT-PCR检测,发现PEG或IMB富集处理提高了阳性样品的检出率。

译名：

Development and application of high sensitivity RT-PCR detection of arabis mosaic virus

作者：

WU Dong-ni;CHEN Shun-sheng;YANG Cui-yun;YU Cui;Shanghai Entry-Exit Inspection and Quarantine Bureau;Shanghai Ocean University;

关键词：

Arabis mosaic virus;;Detection;;RT-PCR;;Polyethylene glycol;;Immunomagnetic beads;;Sensitivity

摘要：

The crude extracts of lily bulbs infected with arabis mosaic virus(ArMV) were concentrated respectively with polyethylene glycol(PEG-6000) and immunomagnetic beads(IMB) and then detected by ELISA,normal RT-PCR and SYBR Green I real-time RT-PCR methods.The results showed that both PEG-and IMB-mediated virus enrichments could greatly improve the sensitivities of those methods to detect ArMV.The PEG-RT-PCR approach could detect the virus from the sample diluted to 10~(-5),and the IMB-RT-PCR approach could detect the virus from the sample diluted to 10~(-3).The real-time RT-PCR detection of ArMV also indicated that the enrichment efficiency of PEG on ArMV was higher than that of IMB.From the RT-PCR detection of ArMV in 19 batches of imported lily bulbs it was found that PEG-and IMB-mediated enrichments could increase the detection rate of positive samples.

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南芥菜花叶病毒高灵敏度检测方法的建立及应用

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