de la Concha-Bermejillo A, Schore C E, Dangler C A, de Mattos C C, de Mattos C A, Osburn B I
Department of Pathology, School of Veterinary Medicine, University of California, Davis 95616.
Am J Vet Res. 1992 Dec;53(12):2245-50.
A slot blot hybridization technique was applied for detection of bluetongue virus (BTV) in blood mononuclear cells (BMNC) obtained from cattle with experimentally induced infection. This technique lacked sensitivity to detect the viral nucleic acid directly in clinical specimens. When aliquots of mononuclear cells from these cattle were cultivated in vitro for 10 days to amplify virus titer, only 33.3% of the samples collected during viremia gave a positive signal in the slot blot hybridization format. By contrast, results for 34.3% of noncultured and 63.3% of cultured mononuclear cell samples collected during viremia were positive by immunofluorescence. The average number of infected cells, as detected by immunofluorescence in the noncultured mononuclear cell samples, was 1 to 5/300,000, and was usually > 10/300,000 in the cultured cell samples. Virus was isolated from all postinoculation blood samples obtained from 4 heifers that were seronegative at the time of inoculation, but was not isolated from any of the preinoculation samples, or from any of the postinoculation samples obtained from 2 heifers that were seropositive at the time of inoculation. When virus isolation was attempted from separated mononuclear cells in 2 heifers, 43.7% of the noncultured and 87.5% of the cultured samples had positive results.
采用斑点杂交技术检测从实验性感染牛获得的血液单核细胞（BMNC）中的蓝舌病病毒（BTV）。该技术缺乏直接检测临床样本中病毒核酸的敏感性。将这些牛的单核细胞等分试样在体外培养10天以扩增病毒滴度，在病毒血症期间采集的样本中，只有33.3% 的样本在斑点杂交检测中呈阳性信号。相比之下，在病毒血症期间采集的未培养单核细胞样本中有34.3% 以及培养单核细胞样本中有63.3% 通过免疫荧光检测呈阳性。通过免疫荧光检测，未培养单核细胞样本中感染细胞的平均数为1至5/300,000，而培养细胞样本中通常>10/300,000。从接种时血清阴性的4头小母牛接种后的所有血液样本中分离到病毒，但未从任何接种前样本或接种时血清阳性的2头小母牛接种后的任何样本中分离到病毒。当尝试从2头小母牛分离的单核细胞中分离病毒时，未培养样本中有43.7% 以及培养样本中有87.5% 得到阳性结果。
