Qi F, Gustad T, Lewis T L, Berry E S
Department of Veterinary & Microbiological Sciences, North Dakota State University, Fargo 58105.
Mol Cell Probes. 1993 Oct;7(5):349-56. doi: 10.1006/mcpr.1993.1052.
A 289 bp cDNA fragment from the 5'-untranslated region (UTR) of 16 bovine viral diarrhoea virus (BVDV) isolates was amplified by reverse transcription and polymerase chain reaction, and sequenced by dideoxy DNA sequencing. The sequence showed greater than 90% homology between the isolates and BVDV NADL in this region, and greater than 97% homology within a 72 base sub-region (nt 314-386). The 289 bp fragment was then used as a probe for rapid detection of BVDV and border disease virus (BDV) from cell culture samples by dot-blot hybridization. This probe hybridized to 100% of BVDV isolates (n = 78) and 100% of BDV isolates (n = 9), but not to the uninfected BT cells or other bovine infectious agents. A shorter probe from the more conserved sub-region also was tested for hybridization with some of the isolates, and the results were similar to those using the longer probe. These results suggest that the 5'-UTR is highly conserved among BVDV and BDV isolates, and may be used as a potential probe for rapid detection of BVDV and BDV in clinical and cell culture samples from cattle and sheep.
通过逆转录和聚合酶链反应扩增了16株牛病毒性腹泻病毒（BVDV）5'-非翻译区（UTR）的一段289 bp的cDNA片段，并采用双脱氧DNA测序法进行测序。该序列在该区域的分离株与BVDV NADL之间显示出大于90%的同源性，在一个72个碱基的子区域（nt 314 - 386）内同源性大于97%。然后，将该289 bp片段用作探针，通过斑点杂交从细胞培养样本中快速检测BVDV和边界病病毒（BDV）。该探针与100%的BVDV分离株（n = 78）和100%的BDV分离株（n = 9）杂交，但不与未感染的BT细胞或其他牛传染性病原体杂交。还测试了来自更保守子区域的较短探针与一些分离株的杂交情况，结果与使用较长探针的结果相似。这些结果表明，5'-UTR在BVDV和BDV分离株中高度保守，可作为快速检测牛羊临床和细胞培养样本中BVDV和BDV的潜在探针。
