A protocol for multiplex PCR assay was established to detect both Banana streak virus (BSV) and Cucumber mosaic virus (CMV) in micropropagated banana cultivars. These viruses are responsible for losses in banana production worldwide. Previous reports have described the integration of BSV in banana genome B. However, the existence of hybrid BSV-free banana cultivars has been reported. Moreover, there have been reports that some CMV strains cannot be mechanically transmitted under laboratory conditions and are not always detected by serological assays. Therefore, the selection of virus-free mother plants for tissue culture is sometimes unsuccessful. The methodology presented in this work overcomes this drawback because it is based on the detection of viral nucleic acids in banana leaf samples. Primers BADNA 1A and BADNA 4 were used to BSV detection, and primers "CMV sense" and "CMV antisense" were adopted to CMV detection. The simultaneous amplification of two different sized amplicons was obtained. Following electrophoresis, a 597 pb fragment correlated with BSV and a 488 fragment correlated with CMV were detected. This assay is an additional tool which should be used to confirm the virus-free status of the mother plants propagated through tissue culture.
Musa spp.; multiplex PCR; BSV; CMV
