Ashbaugh S E, Thompson K E, Belknap E B, Schultheiss P C, Chowdhury S, Collins J K
Veterinary Diagnostic Laboratory, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523, USA.
J Vet Diagn Invest. 1997 Oct;9(4):387-94. doi: 10.1177/104063879700900408.
A sensitive method for simultaneously detecting and discriminating between bovine herpesviruses types 1 and 5 (BHV-1 and BHV-5) was developed using a nested polymerase chain reaction (PCR) technique. Following amplification using type-common primers derived from gC sequences, amplification using type-specific nesting primers produced different-sized bands specific to the corresponding types, as demonstrated by blot hybridization. Less than 0.1 plaque-forming units (PFU) of each virus and 75 fg or less of viral DNA were routinely detected. The PCR technique amplified correct product from 4 BHV-5 isolates and from 48 BHV-1 isolates, all from the United States, and did not amplify heterologous herpesviruses. The PCR technique was more sensitive than virus isolation in detection of BHV-1 or BHV-5 in nasal secretions from experimentally and naturally infected calves, and it detected BHV-1 or BHV-5 in trigeminal ganglia from these calves.
采用巢式聚合酶链反应（PCR）技术，开发了一种同时检测和区分牛1型和5型疱疹病毒（BHV - 1和BHV - 5）的灵敏方法。使用源自gC序列的通用型引物进行扩增后，再使用型特异性巢式引物进行扩增，通过印迹杂交证明会产生对应类型特异的不同大小条带。通常可检测到每种病毒少于0.1个空斑形成单位（PFU）以及75 fg或更少的病毒DNA。PCR技术从4株来自美国的BHV - 5分离株和48株来自美国的BHV - 1分离株中扩增出正确产物，且未扩增出异源疱疹病毒。在检测实验感染和自然感染犊牛鼻分泌物中的BHV - 1或BHV - 5时，PCR技术比病毒分离更灵敏，并且它还能检测到这些犊牛三叉神经节中的BHV - 1或BHV - 5。
