Molecular detection of infectious bursal disease virus by polymerase chain reaction.
The polymerase chain reaction (PCR) technique was applied to the detection of infectious bursal disease virus (IBDV). Reverse transcription followed by the PCR was used to amplify a portion of IBDV genome. A set of primers that specify a 150-base-pair segment of IBDV genome was chosen from an Australian strain of IBDV. Standard challenge strain and variant strains A, D, E, G, and GLS-5 of IBDV serotype 1 and OH strain of serotype 2 from infected bursae were subjected to reverse transcription, followed by 30 cycles of PCR. A single band of the PCR product (DNA) of the expected size from each strain of IBDV was visible on polyacrylamide gels stained with ethidium bromide. Using the same primers, no PCR product was detected from genomic nucleic acids of turkey hemorrhagic enteritis virus, infectious bronchitis virus, reovirus, Salmonella enteritidis, Escherichia coli, and uninfected bursae. The PCR could be efficiently performed on serially diluted IBDV RNA and could detect 2 femtograms of IBDV RNA. The identity of the PCR products was confirmed by direct sequencing. The PCR is a specific and sensitive method for the detection of IBDV.
Wu
C C
CC
College of Veterinary Medicine, Mississippi State University, Mississipi State 39762.
Lin
T L
TL
Zhang
H G
HG
Davis
V S
VS
Boyle
J A
JA
D000818:Animals; D002060:Bursa of Fabricius / Q000382:microbiology*; D004279:DNA, Viral / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D007243:Infectious bursal disease virus / Q000235:genetics / Q000302:isolation & purification*; D015345:Oligonucleotide Probes / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012330:RNA, Double-Stranded / Q000032:analysis*; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D004279:DNA, Viral; D015345:Oligonucleotide Probes; D012330:RNA, Double-Stranded; D012367:RNA, Viral