Detection and differentiation of re-emerging fowlpox virus (FWPV) strains carrying integrated reticuloendotheliosis virus (FWPV-REV) by real-time PCR.
Current strains of fowlpox virus (FWPV) carrying circulating reticuloendotheliosis virus (FWPV-REV) sequence are becoming more pathogenic to poultry. This is evidenced by the fact that vaccination with current available FWPV vaccines provides limited protection against them. To characterize REV insertions in a collection of both older and more recent field isolates, we developed three different types of adjacent oligoprobes and primer sets from specific genomic locations of FWPV and REV: REV-ENV (accession no. K02537, 1382-2260), FWPV-REV integration site (accession no. AF006064, 86-1328), FWPV (accession no. AF198100, 232461-232670), and REV-LTR (accession no. V01204, 305-496). The data indicated that the primers from the REV-ENV region and the TaqMan probes specifically targeted REV-ENV sequences of FWPV-REV strains. Furthermore, the strains were differentiated based on quantitative melting temperature (T(m)) of their amplified products using FRET-based probes. The amplified products were further characterized by sequencing and multiple sequence alignment analysis. The results suggest that integrated REV-ENV sequences are both common and mostly conserved in field isolates. However, the minor variations found within the short-targeted ENV sequence from FWPV-REV strains suggest that these strains could have either undergone periodic point mutational changes or integration with different REV-ENV subtypes.
Tadese
Theodros
T
Department of Pathobiology and Diagnostic Investigation and Diagnostic Center for Population and Animal Health, VMC A-19, Michigan State University, East Lansing, MI 48824, USA. Tadese@dcpah.msu.edu
Fitzgerald
Scott
S
Reed
Willie M
WM
D000818:Animals; D001483:Base Sequence; D052160:Benzothiazoles; D002642:Chick Embryo; D002645:Chickens; D003959:Diamines; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D005587:Fowlpox virus / Q000235:genetics* / Q000302:isolation & purification*; D014644:Genetic Variation; D008969:Molecular Sequence Data; D009930:Organic Chemicals / Q000032:analysis; D016133:Polymerase Chain Reaction; D011804:Quinolines; D017986:Reticuloendotheliosis Viruses, Avian / Q000235:genetics* / Q000302:isolation & purification*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D044366:Transition Temperature; D016662:Virus Integration / Q000235:genetics*
D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I