Three RNA viruses (CYVCV, ICRSV & CTV), one DNA virus (CYMV) and HLB bacterium were detected simultaneously by a multiplex PCR assay.

The primer sets were designed from the conserved genomic regions of viruses for development of mPCR and could be used in both simplex and multiplex PCR.

The multiplex PCR assay developed will help in sensitive and robust detection of prevalent viral and bacterial pathogens of citrus and in production of virus free planting materials and rapid indexing for certification of citrus budwood.

This is the first report on simultaneous detection of Citrus yellow vein clearing virus, Indian citrus ringspot virus, Citrus tristeza virus, Citrus yellow mosaic virus and Huanglongbing bacterium according to our knowledge.