The Banana bunchy top virus (BBTV) and Cucumber mosaic virus (CMV) pose significant threats to banana production, highlighting the need to plant virus-free specimens to maximize yield and control disease spread. Early detection is crucial, as BBTV symptoms typically appear 25 to 85 days after being transmitted by aphids (Pentalonia negronervosa). In this study, we designed multiplex PCR primers targeting key viral regions: movement protein-encoding genes (MV), replication-associated region genes (RP), and coat protein genes (CP) for both BBTV and CMV. Samples were collected from various agroclimatic zones within Maharashtra’s banana-growing regions. An internal control primer was included to identify any PCR reaction failures. Our results show that multiplex PCR is a highly effective method for virus screening in bananas, reducing the occurrence of false positives often seen in single-gene PCR amplification. Based on our findings, we recommend the use of multiplex PCR markers for routine testing of virus-free banana plants to improve disease management and productivity in cavendish banana cultivation.

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