Tissue print hybridisation and reverse transcriptase PCR in the detection of infectious bursal disease viruses in bursal tissues.
The genome segments of infectious bursal disease viruses (IBDV) in the bursa of Fabricius from experimently infected chickens or field samples were detected by tissue print hybridization (TPH) with subsequent reverse transcriptase (RT)- polymerase chain reaction (PCR). Bursae were imprinted onto nylon membrane and then hybridized with a cloned digoxigenin (DIG)-labeled cDNA probe. Tissue prints on nylon membrane were readily distinguished from control prints by color development and differences in signal intensity. In order to verify the TPH test, RT - PCR was used to amplify a 643-base pair fragment on the VP 2 gene of IBDV in the bursa of Fabricius. With all isolates, a c DNA fragment of 643 bp long was generated as expected and further confirmed the specificity of TPH. Our results suggest that a large number of field samples or selected tissues can be rapidly examined by TPH technique when combined with a cloned DIG -labeled c DNA probe.
Liu
H J
HJ
Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan, ROC. hjliu@mail.npust.edu.tw
D000818:Animals; D018175:Birnaviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D002645:Chickens; D017931:DNA Primers; D017403:In Situ Hybridization / Q000379:methods / Q000662:veterinary; D007243:Infectious bursal disease virus / Q000235:genetics / Q000302:isolation & purification; D011201:Poultry Diseases / Q000175:diagnosis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary
D017931:DNA Primers