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Detection of a begomovirus in tomato leaf samples using non-radioactive probes

Major outbreaks of tomato begomoviruses (geminiviruses) in several tomato Lycopersicon esculentum Mill growing in many parts of Brazil have been imposing significant losses upon the tomato agribusiness, due to the introduction of the Bemisia argentifolii (Bemisia tabaci biotype B). Polymerase chain reaction (PCR) and hybridization are generally used for diagnosis. The PCR is a detection method with high sensitivity, however it has the disadvantage of producing false-positives, due to contamination, or false-negatives caused by inhibitors or because of the high primer specifity. The use of nucleic acid hybridization with radio-labelled probes is restricted due to the requirement of special infrastructure and handling experience, the risk for the user’s health and a regular radiochemical element supply. This study is aimed at demonstrating the usefulness of the hybridization method with non-radioactive probes for detection of a tomato begomoviruses in Brazil. The sensitivity of this method was high enabling the detection of 0.1fg of homologous DNA and in crude sap extract diluted up to 100-fold.

Lycopersicon esculentum; geminivirus; diagnosis; digoxigenin


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