【摘要】 发展高效的病毒检测技术已成为当今生物学研究的热点之一。本研究将反转录聚合酶链式反应(PCR)技术与电化学发光分析方法结合起来,用于检测兰花中齿兰环斑病毒。检测过程中采用生物素标记的探针与PCR产物杂交,可起到筛选特异性产物的作用,从而避免假阳性结果的产生。三联吡啶钌标记探针与PCR产物杂交,既可以起到进一步筛选目的片段的作用,又可与分析液中的三丙胺反应,从而产生光信号,被电化学发光仪所检测到。在引物的5′端分别连接一段特异性核酸序列,既提高了钌探针与特异性产物的杂交几率,又增强了样品检测信号,从而提高了信躁比。实验结果表明:此方法灵敏度高、稳定性好、操作简单,可以从兰花样品中准确地检测到齿兰环斑病毒,有望发展成为一种高效的病毒检测方法。更多还原

【Abstract】 It is well known that plant and animal viruses had widely spread the whole of world.It made a big loss in farming and husbandry.The technique for virus detection has become a hot spot in biology research.This research combines reverse transcription polymerase chain reaction(RT-PCR) technique with electrochemilumine-scence(ECL) method to detect Odntoglossum ringspot virus.Biotin-probe hybridizes with PCR product will vesult in selectivity for target detection,soohd pesudo-positive vesult could be avoided.Ru-probe hybridizes with PCR product to emit light for ECL detection.A nucleic acid sequence was linked to 5′ part of the primer,which can improve the chance of hybridization between Ru-probe and PCR product.At the same time,one of the PCR product chain can hybridize two Ru-probe,the ECL signal is intensified.The experiment results show that this method can clearly detect Odntoglossum ring spot virus(ORSV) in orchids.In a word,this method has high sensitivity,stability,simple operation,accurate result,and is a high effective method to detect virus.更多还原