单酶法RTP-CR检测马铃薯纺锤块茎类病毒

Detection of Potato Spindle Tuber Viroid by Single-enzyme RT-PCR

根据Taq酶既有聚合酶活性又有反转录酶活性的特点 ,探索了只在Taq酶单独作用下完成RT PCR反应的条件 ,以达到对马铃薯纺锤块茎类病毒 (PSTV)的快速检测。结果表明 ,反转录阶段退火温度在 37℃、循环数不少于 10次的情况下 ,PCR扩增能得到一条相关特异带。经过嵌合式PCR证明 ,这条带为PSTV的特异带。与其他检测方法相比 ,用单酶法RT PCR检测PSTV步骤简单 。

In single enzyme RT PCR,Taq enzyme plays as roles of both reverse transcripase and DNA polymerase.The reaction conditions of single enzyme RT PCR was researched in this paper in order to detect PSTV rapidly.The results indicated that when the anneal temperature was 37 ℃ and the circles were more than 10 times in reverse transcription phase,a relative band of PSTV was obtained by PCR which could be proved using nested set PCR.Single enzyme RT PCR was simple and low cost,so it could play on important role for the detection of PSTV.