李矮缩病毒RT-PCR方法建立及检测应用

Establishment and Application of Detection of Prune Dwarf Virus (PDV) by RT-PCR

以感病和健康指示植物GF305的总RNA为模板,进行cDNA的合成和PCR扩增,结果从感病材料中扩增出与预期的172bp大小一致的目的片段,而健康的材料无此扩增产物。对此PCR扩增产物克隆测序,进一步佐证了RT-PCR检测结果。经过多次试验验证该检测体系,都可得到很好的重演性,从而建立了李矮缩病毒快速、灵敏、准确的RT-PCR检测技术,并进行了实际应用。

A pair of primers was synthesized based on the nucleotide sequence of the coat protein gene of prune dwarf virus (PDV).The expected size 172bp was amplified by RT-PCR from the infected samples, while no amplified products from the healthy samples.The amplified products were cloned and sequenced. The result showed that the detection system was stable.Thus a rapid, sensitive and accurate method for PDV identification and detection by RT-PCR has been built up and were used to detect whether some fruit trees in orchards were infected by PDV or not.