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However, high cost culture reagents and equipment are required for these methods. As for detection of RBSDV in situ by immunofluorescent labeling, antibodies against RBSDV P7-1, P9-1 and P10 have been adopted for localization of RBSDV in the SBPH midgut. For detection of RBSDV, several methods have been established, such as reverse transcription polymerase chain reaction (RT-PCR), Northern blotting, quantitative real time polymerase chain reaction (qRT-PCR), RT-loop-mediated isothermal DNA amplification (RT-LAMP), single small brown plant hopper RT-PCR and reverse transcription-recombinase polymerase amplification (RT-RPA). The current techniques for RBSDV are based on detection of the gene sequences or viral proteins. It is used as an indicator of RBSDV infection in disease diagnosis. RBSDV P10 protein is the major outer capsid protein. RBSDV is a double-layered icosahedral virus (75–80 nm in diameter) comprised of 10 dsRNA segments (S1–S10) and 6 structural proteins which are P1, P2, P3, P4, P8 and P10 in its viral particle. This is the first report of aptamers being selected for detection of a rice virus capsid protein. These data show that the selected aptamers can be used for the detection of RBSDV P10 protein in vitro and in vivo. All three DNA aptamers can be used to detect RBSDV P10 protein by immunofluorescent labeling in the midgut of RBSDV-infected SBPH. The specificity of these three DNA aptamers was tested through detection of the P10 protein using an enzyme-linked oligonucleotide assay (ELONA) and aptamer-based dot-blot ELISA.

In order to provide a stable and cost-effective detection probe, in this study we selected three DNA aptamers (R3, R5 and R11) by an optimized, standardized and time saving emulsion PCR-based SELEX, for the detection of RBSDV outer-shell P10 protein for in situ localization studies in the midgut of SBPH. RBSDV is transmitted by the small brown planthopper (SBPH, Laodelphax striatellus Fallén) in a persistent manner. Rice black-streaked dwarf virus (RBSDV), classified under the Reoviridae, Fijivirus genus, caused an epidemic in the eastern provinces of China and other East Asian countries and resulted in severe yield loss in rice and wheat production.