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Molecular methods for detection of banana bunchy top virus from banana tissues and viruliferous banana aphids



Molecular methods for detection of banana bunchy top virus from banana tissues and viruliferous banana aphids



Phytopathology 92(6 Supplement): S75, June



The six DNA components of BBTV from Egypt were amplified with PCR primer pairs specific for each component, and the fragments cloned and sequenced (DNA 1, 2, 3, 4, 5, and 6; AF102780, AF102781, AF102782, AF102783, AF102784, AF148139, respectively). Squash and dot blot hybridization methods were developed for detection of BBTV. The probe (480-bp PCR fragment from DNA 1) was labeled with 32P for radioactive detection, with biotin for a chemiluminescent technique, or with digoxigenin for a colorimetric method. The radioactive method was slightly more sensitive than the chemiluminesecent method, whereas the digoxigenin technique was the least sensitive. An immuno-capture PCR (IC-PCR) assay was developed, and it was more sensitive than the hybridization methods. All methods could detect BBTV in extracts of infected banana leaves and in viruliferous banana aphid (Pentalonia negronervosa).

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