Quantitative detection of Cymbidium mosaic virus by real time PCR

作     者：Aichun LIU Yun ZHAO Songlin RUAN Guozheng SHEN 

作者机构：Hangzhou Academy of Agricultural SciencesHangzhou 310024China 

出 版 物：《Frontiers in Biology》 (生物学前沿（英文版）)

年 卷 期：2009年第4卷第3期

页      面：314-320页

核心收录：

学科分类：0202[经济学-应用经济学] 02[经济学] 020205[经济学-产业经济学] 

主　　题：Cymbidium mosaic virus(CyMV) coat protein gene quantitative detection real-time reverse transcription polymerase chain reaction(real-time RTPCR) SYBR Green 

摘      要：The technique of SYBR Green-based quantitative real-time reverse transcription polymerase chain reaction(real-time RT-PCR)was applied to quantitative detect a 764 bp nucleotide sequence containing total coat protein(cp)gene of Cymbidium mosaic virus(CyMV).The plasmid containing the target sequence was constructed to prepare the standard curve and detect the *** standard curve was drawn based on the linear relationship between the logarithm(base 10)of the quantity of target sequence and cycle threshold[C(T)].While the concentration of plasmid DNA falling within the range of 2.6×10^(7)to 2.6×10^(2)copies per tube established a regression equation,y=-0.3583x+10.32,and related coefficient:r^(2)=*** real-time RT-PCR assay for CyMV had a minimum detectable quantity of two copies per *** naturally infected samples of Phalaenopsis *** the artificially inoculated samples of Arachnis *** trace CyMV were quantitatively detected using this *** the positive samples of Phalaenopsis *** Arachnis *** confirmed by DNA sequencing and cp gene homeology *** results showed that CyMV extracted from the leaves of orchid in Hangzhou,Zhejiang Province,China,could be derived from Kunming city(KM),Yunnan Province,*** method characterized by high sensitivity,specificity,and precision is suitable for early diagnosis and quantitative detection of CyMV.