A reliable IC One-step RT-PCR method for the detection of BBrMV to ensure safe exchange of Musa germplasm.
An immunocapture (IC) One-step RT-PCR assay was developed to improve the detection of Banana bract mosaic virus (BBrMV) in single and bulked samples of banana plants. In this paper, an atypical strain of BBrMV was described, the BBrMV "Ref" strain, and we showed that detection with available BBrMV tools using ELISA and RT-PCR approaches was not reliable. Primer sets Bract N1/NR and N2/NR specific to BBrMV were designed and used in RT-PCR and IC-RT-PCR assays with two commercial kits that allow the RT and the PCR reactions to take place simultaneously in the same tube. The new assay enabled detection of BBrMV in leaf extract diluted up to 1 x 10(-10) and in bulked samples of 10 plants, and was proposed as a new international standard to index BBrMV.
Iskra-Caruana
Marie-line
ML
CIRAD-BIOS UMR 385 BGPI, Campus International de Baillarguet, TA A54/K, F-34398 Montpellier Cedex 5, France. marie-line.caruana@cirad.fr
Galzi
Serge
S
Laboureau
Nathalie
N
D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D028521:Musa / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000821:virology*; D017800:Potyvirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D012367:RNA, Viral