One-step RT-PCR for the Detection of Infectious Bursal Disease Virus in Clinical Samples

被引:0
作者
R.S. Kataria
A.K. Tiwari
T. Nanthakumar
P.P. Goswami
机构
[1] National Biotechnology Centre,
[2] Indian Veterinary Research Institute,undefined
来源
Veterinary Research Communications | 2001年 / 25卷
关键词
chickens; contamination; diagnosis; infectious bursal disease; polymerase chain reaction; reverse transcriptase; single-tube technique; sensitivity; specificity; virus;
D O I
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学科分类号
摘要
A single-tube, non-interrupted, one-step RT-PCR has been standardized to amplify the hypervariable region of the VP2 gene sequence of infectious bursal disease virus (IBDV). The technique standardized on purified viral RNA was successfully applied to the detection of the virus directly in clinical samples. The amplified products were confirmed to be IBDV specific by their size in ethidium bromide-stained agarose gel, nested PCR and restriction enzyme digestion. Digestion of the amplicons with StyI restriction enzyme also differentiated classical virus from six very virulent field isolates. The sensitivity of the one-step RT-PCR was found to be 0.2 pg of viral RNA.
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页码:429 / 436
页数:7
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  • [1] Brown M.D.(1994)VP2 sequences of recent European very virulent isolates of infectious bursal disease virus are closely related to each other but are distinct from those of classical strains Journal of General Virology 75 675-680
  • [2] Green P.(1989)Outbreak of virulent infectious bursal disease in East Anglia Veterinary Record 125 271-272
  • [3] Skinner M.A.(1998)Typing of dengue virus in clinical specimens and mosquitoes by single tube multiplex reverse transcriptase PCR Journal of Clinical Microbiology 36 2634-2639
  • [4] Chettle N.(1994)Infectious bursal disease viruses: molecular differentiation of antigenic subtypes among serotype 1 viruses Avian Diseases 38 531-532
  • [5] Stuart J.C.(1987)Antigenic diversity of infectious bursal disease viruses Avian Diseases 31 766-770
  • [6] Wyeth P.J.(1982)Characterization and serological studies of two serotypes of infectious bursal disease virus in turkeys Avian Pathology 26 871-882
  • [7] Harris E.(1997)Detection of bovine herpesvirus-1 (BHV-1) genomic sequence in bovine serum inoculated with BHV-1 by polymerase chain reaction Acta Virologica 41 311-315
  • [8] Roberts T.G.(1998)Detection of infectious bursal disease virus of poultry in clinical samples by RT-PCR Biochemistry and Molecular Biology International 45 315-322
  • [9] Smith L.(1999)Differentiation of infectious bursal disease virus strains by restriction analysis of RT-PCR amplified VP2 gene sequence Acta Virologica 43 243-249
  • [10] Selle J.(2000)Development of probes for differentiation of infectious bursal disease virus strains of various virulence by dot-blot hybridization Acta Virologica 44 259-263