Development and preliminary application of one-step RT-PCR kit for detection of tobacco CMV
To detect tobacco CMV quickly and accurately, a pair of specific primers was designed according to the relatively conservative region in the CMV genomic sequence, and enzymes and buffers needed for reverse transcription and PCR were integrated. By optimizing reaction conditions, a method for detecting CMV-infected tobacco based on one-step RT-PCR was developed and a detection kit was assembled. The specificity, stability and sensitivity of the kit were analyzed. The results showed that the kit featured strong specificity, good repeatability and higher sensitivity. The lowest RNA concentration of template reached 10ng magnitude level. The preliminary application results indicated that the detection results of infected leaves sampled from several areas agreed with the results of ELISA test, the kit could detect CMV successfully.