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双引物探针RT-Realtime PCR检测马铃薯A病毒

作  者:
张京宣;梁炜;耿金培;陈洪俊;杨益娥;粟智平
单  位:
中国检验检疫科学研究院;烟台出入境检验检疫局;山东出入境检验检疫局
关键词:
马铃薯A病毒;CP基因;双引物探针RT-Realtime PCR
摘  要:
马铃薯A病毒(Potato virus A,PVA)是我国重要的检疫性有害生物。本研究根据PVA中CP基因(coat protein gene)的保守序列,设计了两套PCR引物和TaqMan探针,建立了双引物探针RT-RealtimePCR检测PVA的方法。该方法采用实时荧光PCR技术,有效地提高了检测的灵敏度;同时两套引物探针相互验证,有效提高了结果的准确性。实验结果表明,本方法准确、灵敏、简便、快速,检出低限可达0.5fg/μL植物总RNA。
译  名:
Detection of Potato virus A with double primers and probes RT-Realtime-PCR technique
作  者:
Zhang Jingxuan1,Liang Wei1,Geng Jinpei2,Chen Hongjun3,Yang Yi'e2,Su Zhiping2(1.Shangdong Entry–Exit Inspection and Quarantine Bureau,Qingdao 266000,China;2.Yantai Entry–Exit Inspection and Quarantine Bureau;3.Chinese Academy of Inspection and Quarantine)
关键词:
Potato virus A(PVA);coat protein gene;two suits of primers and probes RT-Realtime-PCR
摘  要:
Potato virus A(PVA) is one of the serious quarantine plant pests.Two suits of primers and Taqman probes were designed according to the coat protein gene of PVA in this study,and we set up a new method of RT-Realtime PCR for detection of PVA.The sensitivity was improved effectively by using the realtime PCR technique.At the same time two suits of PCR systems can test each other,which improves effectively the accuracy of the detection results.The method provides a accurate and sensitive detection of PVA,the limit content of the detection is 0.5fg/μL total plant RNA.

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