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Pathobiological investigation of naturally infected canine rabies cases from Sri Lanka.

Abstract The recommended screening of rabies in 'suspect' animal cases involves testing fresh brain tissue. The preservation of fresh tissue however can be difficult under field conditions and formalin fixation provides a simple alternative that may allow a confirmatory diagnosis. The occurrence and location of histopathological changes and immunohistochemical (IHC) labelling for rabies in formalin fixed paraffin embedded (FFPE) canine brain is described in samples from 57 rabies suspect cases from Sri-Lanka. The presence of Negri bodies and immunohistochemical detection of rabies virus antigen were evaluated in the cortex, hippocampus, cerebellum and brainstem. The effect of autolysis and artefactual degeneration of the tissue was also assessed.
PMID
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Authors

Mayor MeshTerms
Keywords

Rabies canine histopathology immunohistochemistry hemi-nested reverse transcription polymerase chain reaction

Journal Title bmc veterinary research
Publication Year Start




PMID- 28403882
OWN - NLM
STAT- MEDLINE
DA  - 20170413
DCOM- 20170418
LR  - 20170418
IS  - 1746-6148 (Electronic)
IS  - 1746-6148 (Linking)
VI  - 13
IP  - 1
DP  - 2017 Apr 12
TI  - Pathobiological investigation of naturally infected canine rabies cases from Sri 
      Lanka.
PG  - 99
LID - 10.1186/s12917-017-1024-5 [doi]
AB  - BACKGROUND: The recommended screening of rabies in 'suspect' animal cases
      involves testing fresh brain tissue. The preservation of fresh tissue however can
      be difficult under field conditions and formalin fixation provides a simple
      alternative that may allow a confirmatory diagnosis. The occurrence and location 
      of histopathological changes and immunohistochemical (IHC) labelling for rabies
      in formalin fixed paraffin embedded (FFPE) canine brain is described in samples
      from 57 rabies suspect cases from Sri-Lanka. The presence of Negri bodies and
      immunohistochemical detection of rabies virus antigen were evaluated in the
      cortex, hippocampus, cerebellum and brainstem. The effect of autolysis and
      artefactual degeneration of the tissue was also assessed. RESULTS: Rabies was
      confirmed in 53 of 57 (93%) cases by IHC. IHC labelling was statistically more
      abundant in the brainstem. Negri bodies were observed in 32 of 53 (60.4%) of the 
      positive cases. Although tissue degradation had no effect on IHC diagnosis, it
      was associated with an inability to detect Negri bodies. In 13 cases, a
      confirmatory Polymerase chain reaction (PCR) testing for rabies virus RNA was
      undertaken by extracting RNA from fresh frozen tissue, and also attempted using
      FFPE samples. PCR detection using fresh frozen samples was in agreement with the 
      IHC results. The PCR method from FFPE tissues was suitable for control material
      but unsuccessful in our field cases. CONCLUSIONS: Histopathological examination
      of the brain is essential to define the differential diagnoses of behaviour
      modifying conditions in rabies virus negative cases, but it is unreliable as the 
      sole method for rabies diagnosis, particularly where artefactual change has
      occurred. Formalin fixation and paraffin embedding does not prevent detection of 
      rabies virus via IHC labelling even where artefactual degeneration has occurred. 
      This could represent a pragmatic secondary assay for rabies diagnosis in the
      field because formalin fixation can prevent sample degeneration. The brain stem
      was shown to be the site with most viral immunoreactivity; supporting recommended
      sampling protocols in favour of improved necropsy safety in the field. PCR
      testing of formalin fixed tissue may be successful in certain circumstances as an
      alternative test.
FAU - Beck, S
AU  - Beck S
AD  - Pathology Department, Animal and Plant Health Agency, Weybridge, UK.
      [email protected]
FAU - Gunawardena, P
AU  - Gunawardena P
AD  - Department of Veterinary Pathobiology, University of Peradeniya, Peradeniya, Sri 
      Lanka.
FAU - Horton, D L
AU  - Horton DL
AD  - Wildlife Zoonoses and Vector Borne Diseases Research Group, Animal and Plant
      Health Agency, Weybridge, UK.
FAU - Hicks, D J
AU  - Hicks DJ
AD  - Pathology Department, Animal and Plant Health Agency, Weybridge, UK.
FAU - Marston, D A
AU  - Marston DA
AD  - Wildlife Zoonoses and Vector Borne Diseases Research Group, Animal and Plant
      Health Agency, Weybridge, UK.
FAU - Ortiz-Pelaez, A
AU  - Ortiz-Pelaez A
AD  - Animal and Plant Health Agency, Weybridge, UK.
FAU - Fooks, A R
AU  - Fooks AR
AD  - Wildlife Zoonoses and Vector Borne Diseases Research Group, Animal and Plant
      Health Agency, Weybridge, UK.
FAU - Nunez, A
AU  - Nunez A
AD  - Pathology Department, Animal and Plant Health Agency, Weybridge, UK.
LA  - eng
PT  - Journal Article
DEP - 20170412
PL  - England
TA  - BMC Vet Res
JT  - BMC veterinary research
JID - 101249759
RN  - 0 (Antigens, Viral)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Animals
MH  - Antigens, Viral/analysis
MH  - Brain/pathology/virology
MH  - Dog Diseases/diagnosis/*pathology
MH  - Dogs
MH  - Polymerase Chain Reaction
MH  - RNA, Viral/genetics
MH  - Rabies/*diagnosis/genetics/pathology/*veterinary
MH  - Sri Lanka
MH  - Tissue Fixation/standards/veterinary
PMC - PMC5389160
OTO - NOTNLM
OT  - Rabies canine histopathology immunohistochemistry hemi-nested reverse
      transcription polymerase chain reaction
EDAT- 2017/04/14 06:00
MHDA- 2017/04/19 06:00
CRDT- 2017/04/14 06:00
PHST- 2016/02/03 [received]
PHST- 2017/04/03 [accepted]
AID - 10.1186/s12917-017-1024-5 [doi]
AID - 10.1186/s12917-017-1024-5 [pii]
PST - epublish
SO  - BMC Vet Res. 2017 Apr 12;13(1):99. doi: 10.1186/s12917-017-1024-5.

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