Simultaneously detection of three bean viruses, Bean common mosaic virus, Bean common mosaic necrosis virus and Cucumber mosaic virus by Multiplex-RT-PCR*

Bean common mosaic virus (BCMV), Bean common mosaic necrosis virus (BCMNV) and Cucumber mosaic virus (CMV) are seed-borne viruses in beans causing severe damage and reduction of yield and quality of this crop. Since there are lots of reports on the occurrence of mixed infection of BCMV, BCMNV and CMV, thus setting up an accurate, reliable and rapid method for detecting of these viruses in bean crops could play an important role in prevention of their spread and damages and also screening of healthy plants. For this purpose, in the present study, we tried to set up a multiplex RT-PCR for simultaneous detection of BCMV, BCMNV and CMV as well as an internal control gene, 18S rRNA. To do this end, the infection of bean plants to above mentioned viruses was confirmed by DAC-ELISA. Following total RNA extraction, cDNA was synthesized using random hexamer primers. cDNA fragments were successfully amplified using specific primers in expected sizes as almost 470 bp for BCMV, 500bp for internal control, 704 bp for BCMNV and 870 bp for CMV, respectively. The results of this study showed that this method is capable of detecting BCMV, BCMNV and CMV in 100 ng of total RNA.