Venter E H, Huismans H, Van Dijk A A
Department of Veterinary Tropical Diseases, University of Pretoria, Onderstepoort, South Africa.
Onderstepoort J Vet Res. 1995 Dec;62(4):217-22.
The serogroup specificity of the bluetongue virus (BTV) NS1 and VP3 gene probes was confirmed by means of northern blot hybridization. Under high-stringency conditions both probes hybridized to 22 BTV serotypes (18 South African serotypes, BTV3 from Cyprus and BTV16 from Pakistan) but not to serotypes that originate from Australia and India. Furthermore, NS1 gene probes of BTV and African horsesickness virus (AHSV) were used in a dot-spot in situ hybridization procedure to differentiate between BTV and AHSV in co-infected cell cultures. The method detects viral RNA directly i glutaraldehyde-fixed infected cell cultures without prior nucleic-acid extraction or purification. AHSV could be detected in cells infected with AHSV at a multiplicity of infection of 10(-4) PFU/cell in the presence of a hundred excess of co-infecting BTV. The method may have an application in epidemiological surveys to detect different orbiviruses in the same Culicoides population.
通过Northern印迹杂交法证实了蓝舌病毒（BTV）NS1和VP3基因探针的血清群特异性。在高严格条件下，两种探针均与22种BTV血清型（18种南非血清型、来自塞浦路斯的BTV3和来自巴基斯坦的BTV16）杂交，但不与源自澳大利亚和印度的血清型杂交。此外，BTV和非洲马瘟病毒（AHSV）的NS1基因探针用于斑点原位杂交程序，以区分共感染细胞培养物中的BTV和AHSV。该方法可直接在戊二醛固定的感染细胞培养物中检测病毒RNA，无需事先进行核酸提取或纯化。在存在一百倍过量共感染BTV的情况下，能够在感染复数为10^(-4) PFU/细胞的AHSV感染细胞中检测到AHSV。该方法可能在流行病学调查中用于检测同一库蠓种群中的不同环状病毒。
