Detection of avian leukosis virus subgroup J using the polymerase chain reaction.
A polymerase chain reaction (PCR) assay was developed for the detection of avian leukosis virus strain J (ALV-J) in chickens. Primers were based in the E element and the 3' terminus of the long terminal repeat of proviral ALV-J. PCR products were amplified from genomic DNA extracted from chicken embryo fibroblasts (CEF) infected with either strain HPRS-103, the prototype of ALV-J, or field isolates of ALV-J obtained from broiler breeder flocks in the United States that exhibited myeloid leukosis. The newly developed PCR detected ALV-J in DNA prepared from CEF inoculated with ALV-J but not from CEF inoculated with subgroup A, B, C, D, or E. The PCR also detected ALV-J in DNA prepared from blood, combs, and toes obtained from chickens experimentally infected with ALV-J and in DNA obtained from peripheral blood monocytes from naturally infected broiler breeder chickens. The PCR described here offers a specific and sensitive alternative to conventional virus isolation tests for ALV-J.
Smith
E J
EJ
U.S. Department of Agriculture, Agricultural Research Service, Avian Disease and Oncology Laboratory, Lansing, MI 48823, USA.
Williams
S M
SM
Fadly
A M
AM
D000818:Animals; D001353:Avian Leukosis / Q000175:diagnosis*; D001354:Avian Leukosis Virus / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D002478:Cells, Cultured / Q000382:microbiology; D002642:Chick Embryo; D002645:Chickens / Q000382:microbiology*; D003001:Cloning, Molecular / Q000379:methods; D004279:DNA, Viral / Q000032:analysis* / Q000737:chemistry; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity; D017421:Sequence Analysis / Q000379:methods / Q000662:veterinary; D012689:Sequence Homology, Nucleic Acid
D004279:DNA, Viral