Background: Chrysanthemum plants are subject to serious virus diseases. The viruses cause severe losses of quantity and quality of chrysanthemum. The most problematic pathogens of chrysanthemum are typically considered to be Chrysanthemum virus B (CVB). A method for the simultaneous detection of Chrysanthemum virus B is needed.

Results: We use specific primers, which is derived from coat protein region, for reverse transcription to obtain cDNA. At the same time, the nested amplification PCR is employed to detect it. The sensitivity is up to the 10^-9, greatly improved the sensitivity.

Conclusion: A highly specific and sensitive nested PCR-based assay is described for detecting Chrysanthemum virus B. This new method is highly specific and sensitive for detection of Chrysanthemum virus B, which are known to infect chrysanthemum plants in the field. Further, this protocol has an advantage over traditional methods in that it is more cost-effective.  This assay is ideal for performing an early stage diagnosis of the disease.