Specific detection of shedding and latency of bovine herpesvirus 1 and 5 using a nested polymerase chain reaction

被引:61
作者
Ashbaugh, SE [1 ]
Thompson, KE
Belknap, EB
Schultheiss, PC
Chowdhury, S
Collins, JK
机构
[1] Colorado State Univ, Coll Vet Med & Biomed Sci, Vet Diagnost Lab, Ft Collins, CO 80523 USA
[2] Colorado State Univ, Coll Vet Med & Biomed Sci, Dept Clin Sci, Ft Collins, CO 80523 USA
[3] Colorado State Univ, Coll Vet Med & Biomed Sci, Dept Pathol, Ft Collins, CO 80523 USA
[4] Colorado State Univ, Coll Vet Med & Biomed Sci, Dept Microbiol, Ft Collins, CO 80523 USA
[5] Kansas State Univ, Coll Vet Med, Dept Pathol & Microbiol, Manhattan, KS 66506 USA
关键词
D O I
10.1177/104063879700900408
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
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.
引用
收藏
页码:387 / 394
页数:8
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