Development of a shuttle polymerase chain reaction for the detection of bovine herpesvirus 2.
Three different polymerase chain reaction (PCR) protocols were evaluated for their ability to detect bovine herpesvirus 2 (BoHV-2): single-step PCR with 3 reaction stages (denaturation, annealing and extension), 2 reaction stages (denaturation and annealing/extension; shuttle PCR), and semi-nested PCR with 3 reaction stages. All the PCR protocols showed the same sensitivity (detection limit of 0.4 TCID(50)). A non-specific band sometimes appeared in mock cell DNA at annealing temperatures below 64 degrees C. The shuttle PCR was found to be superior to the other protocols under consideration because of the speed of its application. Furthermore, no non-specific band was detected in DNAs of eight other DNA viruses. Thus, the shuttle PCR seems to be an excellent diagnostic tool for BoHV-2 infections.
Imai
Kunitoshi
K
Hokkaido Research Station, National Institute of Animal Health, Sapporo, Hokkaido, Japan.
Ishihara
Ryoko
R
Jayawardane
Gamini W
GW
Nishimori
Kei
K
Nishimori
Tomoko
T
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000662:veterinary; D001911:Herpesvirus 2, Bovine / Q000235:genetics* / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004279:DNA, Viral