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RT-PCR检测齿兰环斑病毒技术的建立

Development of RT-PCR assay for the detection of Odntoglossum rongspot virus

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【作者】 明艳林李梅郑国华吴祖建

【Author】 MING Yanlin1, LI Mei1, ZHENG Guohua2, WU Zujian2 (1.National Plant Introduction Quarantine Base, Xiamen Overseas Chinese Subtropical Plant Introduction Garden, Xiamen, Fujian 361002, China;2.Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China)

【机构】 厦门华侨亚热带植物引种园国家植物引种隔离检疫基地福建农林大学植物病毒研究所福建农林大学植物病毒研究所 福建厦门361002福建厦门361002福建福州 3 50 0 0 2福建福州 3 50 0 0 2

【摘要】 根据已报道的齿兰环斑病毒(ORSV)外壳蛋白(CP)基因序列,进行同源性比较,设计了一对各为20bp的引物.通过优化试验条件,建立了稳定的RT-PCR检测ORSV体系,其检测灵敏度可达0.01ng·mL-1.将此方法与ELISA进行比较,结果表明,RT-PCR检测灵敏度比ELISA高1000倍.应用这两种方法同时检测了138瓶兰花组培苗样品,其中RT-PCR检测出32瓶兰花组培苗感染病毒,而ELISA只能检测其中的18瓶.

【Abstract】 Using a pair of 20 bp primers complementary to the conserved Odntoglossum ringspot virus (ORSV) coat protein (CP) gene sequences published previously, the sensitive RTPCR method was set up to test ORSV immediately by optimizing experiment conditions. The sensitivity of RTPCR method can come to 0.01 ng·mL-1 ORSV, which is 1000 times higher than ELISA. And 138 samples suspected to be infected with ORSV were tested by both of the two methods. The results showed that 32 of the samples were positive by RTPCR, but only 18 were positive by ELISA.

【关键词】 齿兰环斑病毒RT-PCR检测
【Key words】 Odntoglossum ringspot virusRT-PCRdetection
【基金】 福建省自然科学基金资助项目(B9910027).
  • 【文献出处】 福建农业大学学报 , 编辑部邮箱 ,2003年03期
  • 【分类号】S436.8
  • 【被引频次】26
  • 【下载频次】169
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