Rolling Circle Amplification Is More Sensitive than PCR and Serology-Based Methods in Detection of Banana streak virus in Musa Germplasm

Abstract

Banana (Musa sp.) is a popular and important crop among many communities in East Africa. Banana production is however threatened by the wide-spread banana streak disease (BSD), caused by Banana streak virus (BSV). The suc- cess of BSV management is inherently coupled to the availability of a sensitive indexing method. In this study, the sen- sitivity of three BSV detection techniques: rolling circle amplification (RCA), immunocapture PCR (with degenerate and Gold finger primers) and standard PCR was compared. A set of 32 BSD-asymptomatic samples were used to com- pare the techniques. Analysis of variance (ANOVA) for comparison of the four techniques showed that there were sig- nificant differences (P < 0.05) among all the means, with RCA and direct PCR having the highest detection mean values. Owing to its fidelity and capacity to circumvent the amplification of the integrated nuclear viral sequences, the RCA technique is recommended for routine indexing of Musa tissues for BSV. This study unveils a more reliable BSV detec- tion method, a need that has remained unaddressed for a long while.