Naif H M, Brandon R B, Daniel R C, Lavin M F
Queensland Institute of Medical Research, Herston, Brisbane, Australia.
Vet Microbiol. 1990 Nov;25(2-3):117-29. doi: 10.1016/0378-1135(90)90071-3.
Bovine leukaemia virus (BLV) is the causative agent in enzootic bovine leukosis a disease occurring worldwide. This virus is normally detected by the agar gel immunodiffusion or ELISA assays which rely on the appearance of antibodies to a major surface protein of the virus, gp51, present in the serum of infected cattle. We have used the polymerase chain reaction, which depends on the amplification of specific DNA sequences as a sensitive assay for the detection of BLV. It was possible to detect proviral DNA in 100 pg of tumour DNA from an infected host using agarose gel electrophoresis followed by ethidium bromide staining. The sensitivity of the assay was increased by two log orders when hybridization analysis, using a BLV proviral DNA probe, was used in combination with amplification of the DNA. Proviral DNA was detected in both lymphocytic and tumour DNA and at all stages of infection in cattle.
牛白血病病毒(BLV)是地方流行性牛白血病的病原体,这种疾病在全球范围内都有发生。该病毒通常通过琼脂凝胶免疫扩散或ELISA检测法进行检测,这些方法依赖于感染牛血清中针对病毒主要表面蛋白gp51的抗体的出现。我们使用了聚合酶链反应,该反应依赖于特定DNA序列的扩增,作为检测BLV的灵敏检测方法。使用琼脂糖凝胶电泳,随后用溴化乙锭染色,能够从感染宿主的100 pg肿瘤DNA中检测到前病毒DNA。当使用BLV前病毒DNA探针进行杂交分析并与DNA扩增相结合时,检测方法的灵敏度提高了两个数量级。在牛的淋巴细胞和肿瘤DNA以及感染的各个阶段都检测到了前病毒DNA。