Genotypic screening of pseudorabies virus strains for thymidine kinase deletions by use of the polymerase chain reaction.
Genetic recombination between field strains and vaccine strains of pseudorabies virus (PRV) has been suggested as a scenario that might arise from use of deletion-mutant modified-live vaccine strains, particularly those strains attenuated by deletions within the thymidine kinase (TK-) gene locus. To address this hypothesis experimentally, it is necessary to screen large numbers of PRV isolates for their TK genotype. Techniques to detect the native TK genotype are routinely used in molecular virology laboratories, but are time-consuming. We adapted the polymerase chain reaction to define the genotypic status of PRV isolates with regard to the presence or absence of deletions in the TK gene locus. Used in tandem with the existing glycoprotein-specific ELISA that discriminate between PRV-vaccinated and field strain-infected swine populations, the described technique may help to clarify whether vaccine-derived recombinants are generated under natural conditions and after normal vaccine usage.
Dangler
C A
CA
Department of Veterinary Science, College of Agriculture, Pennsylvania State University, University Park 16802.
Deaver
R E
RE
Kolodziej
C M
CM
Rupprecht
J D
JD
D000818:Animals; D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis* / Q000737:chemistry; D015252:Deoxyribonucleases, Type II Site-Specific; D005838:Genotype; D011558:Herpesvirus 1, Suid / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D013937:Thymidine Kinase / Q000235:genetics*; D014709:Vero Cells
D004279:DNA, Viral; D013937:Thymidine Kinase; C025651:CCCGGG-specific type II deoxyribonucleases; C034510:CTCGAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific