Detection of infectious bursal disease virus infection using the polymerase chain reaction.
The method of reverse transcription (RT) followed by the polymerase chain reaction (PCR) was used to amplify two different fragments of the infectious bursal disease virus (IBDV) genome. Two sets of primer framed two different regions within the genes coding for proteins VP2 and VP3, respectively. Both sequences were detected in five strains of IBDV, whereas, none were obtained from uninfected control cells. The sensitivity of RT-PCR was carried out on nucleic acids from the IBDV infected cell cultures. The detection limit was 10(0) to 10(-1) TCID50 in ethidium bromide stained gels and could be enhanced further to 10(-1) to 10(-3) TCID50 by hybridization after southern transfer. In addition, detection of IBDV infection in 12 out of 14 bursal specimens examined by this technique was shown to be entirely consistent with the clinical history and an alternative diagnostic method. The speed, sensitivity, and specificity of this method is relevant for the diagnosis of infection with IBDV.
Lee
L H
LH
Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan, People's Republic of China.
Yu
S L
SL
Shieh
H K
HK
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D015342:DNA Probes; D004279:DNA, Viral / Q000235:genetics; D005069:Evaluation Studies as Topic; D007243:Infectious bursal disease virus / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011201:Poultry Diseases / Q000175:diagnosis; D012088:Reoviridae Infections / Q000175:diagnosis / Q000662:veterinary; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
D015342:DNA Probes; D004279:DNA, Viral