南芥菜花叶病毒高灵敏度检测方法的建立及应用  

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

作　　者：吴东妮[1,2] 陈舜胜[2] 杨翠云[1] 于翠[1] 

机构地区：[1]上海出入境检验检疫局,上海200135 [2]上海海洋大学,上海201306

出　　处：《上海农业学报》2013年第5期74-78,共5页Acta Agriculturae Shanghai

基　　金：国家质检总局科技项目(2011IK172)资助

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。The crude extracts of lily bulbs infected with arabis mosaic virus （ArMV）were concen- trated 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 di- luted to 10-s ,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.

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 

分 类 号：S436.44[农业科学—农业昆虫与害虫防治]