Specific detection of shedding and latency of bovine herpesvirus 1 and 5 using a nested polymerase chain reaction.
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.
Ashbaugh
S E
SE
Veterinary Diagnostic Laboratory, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523, USA.
Thompson
K E
KE
Belknap
E B
EB
Schultheiss
P C
PC
Chowdhury
S
S
Collins
J K
JK
D018138:Alphaherpesvirinae / Q000302:isolation & purification* / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases; D002460:Cell Line; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D007242:Herpesvirus 1, Bovine / Q000302:isolation & purification* / Q000502:physiology; D008168:Lung; D009297:Nasal Mucosa / Q000378:metabolism / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D012680:Sensitivity and Specificity; D010948:Viral Plaque Assay; D017735:Virus Latency; D017201:Virus Shedding
D017931:DNA Primers; D004279:DNA, Viral