Development of improved molecular methods for the detection of deformed wing virus (DWV) in honeybees (Apis mellifera L.) and mites ( Varroa destructor Oud.).
A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful either as template for RT-PCR or in nucleic acids hybridization, was developed. Sensitivity of the methods were evaluated up to 10(9) and 10(6) dilution of TNAs extracted from a single honeybee, for reverse transcriptase polymerase chain reaction and molecular hybridization respectively. The two diagnostic methods developed could be useful for the study of the molecular biology and the pathology of DWV. For practical applications dot-blot hybridization could be used in order to study the incidence of DWV in honeybees populations. The method is enough sensitive, rapid and less affected by contamination problems compared to RT-PCR and thus it could be applied to the sanitary certification of honeybees and their products.
Parrella
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G
Istituto per la Protezione delle Piante del CNR Via Università 133, 80055 Portici (NA), Italy. giuseppe.parrella@ipp.cnr.it
Caprio
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E
Mazzone
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P
D000818:Animals; D001516:Bees / Q000821:virology*; D006824:Hybridization, Genetic; D015151:Immunoblotting; D007304:Insect Viruses / Q000235:genetics / Q000302:isolation & purification*; D008925:Mites / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D012367:RNA, Viral