Detection of bluetounge virus serotype 17 in Culicoides variipennis by nucleic acid blot and sandwich hybridization techniques

Schoepp, R.J.; Bray, J.F.; Olson, K.E.; El-Hussein, A.; Holbrook, F.R.; Blair, C.D.; Roy, P.; Beaty, B.J.

Detection of bluetounge virus serotype 17 in Culicoides variipennis by nucleic acid blot and sandwich hybridization techniques

1990

Molecular hybridization techniques were developed for the detection and surveillance of bluetongue virus (BTV) serotype 17 in the insect vector Culicoides variipennis, abiting midge. Radiolabeled RNA and cDNA probes were generated from sequences of the L3 segment of BTVserotype 17. These probes were used to detect BTV RNA in pools of infected C. variipennis by hybridizing theprobes directly to analyte immobilized on nylon membranes or by using a nucleic acid sandwich hybridization test. Hybridization procedures were able to detect 1 infected C. variipennis in a pool of 50 and as little as3.55 log10 50% tissue culture infective doses per ml of virus. These hybridization techniques provide an alternativeto virus isolation for the surveillance of BTV in vector populations.

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