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

James S. Guy, Melissa A. West, Frederick J. Fuller, Rosemary A. Marusak, H L Shivaprasad, James L. Davis, Oscar J. Fletcher

Research output: Contribution to journalArticle

7 Citations (Scopus)

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 consistent with 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.

Original languageEnglish (US)
Pages (from-to)70-75
Number of pages6
JournalAvian Diseases
Volume55
Issue number1
DOIs
StatePublished - Mar 2011

Fingerprint

proventriculitis
Reverse Transcriptase Polymerase Chain Reaction
Chickens
necrosis
Necrosis
Proventriculus
reverse transcriptase polymerase chain reaction
chickens
Viruses
viruses
proventriculus
lesions (animal)
formalin
Paraffin
alkanes
Formaldehyde

Keywords

  • chicken
  • polymerase chain reaction
  • proventriculitis

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Food Animals
  • Immunology and Microbiology(all)

Cite this

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. / Guy, James S.; West, Melissa A.; Fuller, Frederick J.; Marusak, Rosemary A.; Shivaprasad, H L; Davis, James L.; Fletcher, Oscar J.

In: Avian Diseases, Vol. 55, No. 1, 03.2011, p. 70-75.

Research output: Contribution to journalArticle

Guy, James S. ; West, Melissa A. ; Fuller, Frederick J. ; Marusak, Rosemary A. ; Shivaprasad, H L ; Davis, James L. ; Fletcher, Oscar J. / 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. In: Avian Diseases. 2011 ; Vol. 55, No. 1. pp. 70-75.
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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 consistent with 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.",
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