title: Detection of chicken proventricular necrosis virus (R11/3 virus) in experimental and naturally occurring cases of transmissible viral proventriculitis with the use of a reverse transcriptase-PCR procedure.
abstract: A reverse-transcriptase-polymerase-chain-reaction (RT-PCR) procedure was evaluated for detection of chicken proventricular necrosis virus (CPNV) in transmissible viral proventriculitis (TVP) -affected chickens. The RT-PCR procedure was compared with indirect immunofluorescence (IFA) and virus isolation for detection of CPNV in experimentally infected chickens. Microscopic lesions characteristic of TVP were detected on days 5-35 postexposure (PE) in CPNV-infected chickens; CPNV was detected by RT-PCR on days 3-14 PE in freshly collected proventriculi, and on days 1-14 PE in formalin-fixed paraffin-embedded (FFPE) proventriculi. CPNV was detected in proventriculi of experimentally infected chickens by IFA on days 3-10 PE, and by virus isolation on days 1-14 PE. With IFA used as a reference, sensitivity of the RT-PCR procedure with freshly collected and FFPE proventriculi was 88% and 100%, respectively; specificity was 83% and 86%, respectively. Proventriculi (FFPE) obtained from suspect TVP cases (n=19) were evaluated for presence of CPNV by RT-PCR and microscopic lesions consistentwith TVP. CPNV was detected by RT-PCR in proventriculi from 8/11 TVP (+) cases (24/36 tissue sections). TVP (+) cases were defined by microscopic lesions characteristic of TVP; CPNV was not detected in proventriculi (0/8 cases, 0/32 tissue sections) in the absence of these lesions. The association between presence of TVP-characteristic microscopic lesions and presence of CPNV was highly significant (P = 0.0014). These findings indicate the utility of the RT-PCR procedure for detection of CPNV and provide additional evidence for an etiologic role for this virus in TVP.
authors.affiliation: Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina 27606, USA. jim_guy@ncsu.edu
authors.forename: James S
authors.initials: JS
authors.lastname: Guy
authors.forename: Melissa A
authors.initials: MA
authors.lastname: West
authors.forename: Frederick J
authors.initials: FJ
authors.lastname: Fuller
authors.forename: Rosemary A
authors.initials: RA
authors.lastname: Marusak
authors.forename: H L
authors.initials: HL
authors.lastname: Shivaprasad
authors.forename: James L
authors.initials: JL
authors.lastname: Davis
authors.forename: Oscar J
authors.initials: OJ
authors.lastname: Fletcher
mesh_terms: D000257:Adenoviridae Infections / Q000662:veterinary*; D000818:Animals; D000259:Aviadenovirus; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D011201:Poultry Diseases / Q000821:virology*; D011531:Proventriculus / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*