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一种豇豆重花叶病毒RT-PCR检测方法的研究

作  者:
李彬;吴翠萍;粟寒;安榆林
单  位:
江苏出入境检验检疫局
关键词:
豇豆重花叶病毒;简并引物;反转录PCR;双抗夹心酶联免疫法
摘  要:
对一种豇豆重花叶病毒(Cowpeas evere mosaic virus,CPSMV)反转录PCR分子检测方法进行了研究。利用从美国菌种保藏中心和德国微生物毒株保藏中心引进的CPSMV标准毒株,设计了一对特异性简并引物(CPSMVf/CPSMVr),建立了CPSMVRT-PCR检测方法。该方法具有良好的扩增反应及特异性,灵敏度可达1pg的总病叶组织RNA量。CPSMV双抗夹心酶联免疫法(DAS-ELISA)灵敏度为10μg叶组织。该方法适用于豇豆等豆类蔬菜种子及种苗中对豇豆重花叶病毒的快速检测。
译  名:
Research on a method of RT-PCR for detection of Cowpea severe mosaic virus
作  者:
Li Bin,Wu Cuiping,Su Han,An Yulin(Jiangsu Entry-Exit Inspection and Quarantine Bureau,Nanjing 210001,China)
关键词:
CPSMV;degenerate primer;RT -PCR;DAS-ELISA
摘  要:
A method of reverse transcript(RT)PCR was researched for detection of Cowpea severe mosaic virus (CPSMV). By using standard CPSMV strains introduced from ATCC and DSMZ as materials,RT-PCR method which showed good amplification reaction and specificity was constituted and its sensitivity could reach 1 pg total RNA of leaf tissue infected by CPSMV. Meanwhile,the sensitivity of double antibody sandwich ELISA (DAS-ELISA) for test of CPSMV was only 10μg leaf tissue. The RT-PCR method was suitable for rapid detection of CPSMV from legume seeds and seedlings such as cowpea.

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