DETECTION OF BLUETONGUE VIRUS IN CLINICAL-SAMPLES BY POLYMERASE CHAIN-REACTION

A previously described bluetongue virus (BTV) serogroup polymerase cha in reaction (PCR) assay was applied to clinical samples. The sensitivi ty of the BTV serogroup PCR was increased by the use of non-radioactiv e chemiluminescent hybridization. Unfractionated whole blood samples f rom rams experimentally inoculated with cell culture-adapted BTV-11 UC -8 were analyzed by virus isolation (VI) on Vero cells and PCR. VI and PCR were in agreement, with the exception of 3 blood samples that wer e VI negative and PCR positive. In semen spiked with BTV-11 UC-8, PCR detected as little as 1.6 x 10(2) plaque-forming units of BTV/ml of se men. BTV in the spleen of a sheep submitted for necropsy for suspect B TV infection was detected by both PCR and VI in embryonated chicken eg gs. BTV PCR with nonradioactive chemiluminescent hybridization resulte d in a level of sensitivity comparable to that of VI and likly more se nsitive than VI on Vero cells for blood. This BTV PCR has great promis e for rapid, sensitive, and specific detection of active BTV infection in a variety of clinical samples.