用各种类型的DNA探针检测牛疱疹病毒1型。

Detection of bovine herpesvirus 1 with various types of DNA probes.

作者信息

Vilcek S, Deliová I, Forgác O, Strojný L, Takácsová I, Harvan M, Benko G

机构信息

Laboratory of Genetic Manipulations, Institute of Experimental Veterinary Medicine, Slovakia.

出版信息

Acta Vet Hung. 1993;41(1-2):179-90.

PMID:8116497

Abstract

The method of dot-blot hybridization on nitrocellulose filters by various types of DNA probes (ds recombinant plasmids, ss recombinant M13 phages and a 42bp synthetic oligonucleotide) was used for BHV-1 detection. The highest sensitivity was achieved with the 32P-pUR1 probe (1.8 kb random EcoRI-HindIII fragment inserted into pUC9) which detected the BHV-1 genome in 5 x 10(3) infected MDBK cells. Using the pUR1 probe, no cross hybridization was observed with other herpesviruses: BHV-2, 3, 4, and Aujeszky's disease virus. The 32P-pUR1 probe detected BHV-1 in nasal swabs of calves as early as on day 1 after experimental infection. The maximum intensity of BHV-1 detection occurred on day 1-3. The 32P-pUR1 probe also detected BHV-1 in field samples of nasal swabs from cows and calves.

摘要

采用多种类型的DNA探针（双链重组质粒、单链重组M13噬菌体和42bp合成寡核苷酸）在硝酸纤维素滤膜上进行斑点杂交的方法来检测牛疱疹病毒1型（BHV-1）。32P-pUR1探针（插入pUC9的1.8kb随机EcoRI-HindIII片段）具有最高的灵敏度，能在5×10³个感染的MDBK细胞中检测到BHV-1基因组。使用pUR1探针时，未观察到与其他疱疹病毒（BHV-2、3、4和奥耶斯基氏病病毒）的交叉杂交。32P-pUR1探针在实验感染后第1天就能在犊牛鼻拭子中检测到BHV-1。检测BHV-1的最大强度出现在第1至3天。32P-pUR1探针还能在奶牛和犊牛鼻拭子的现场样本中检测到BHV-1。