Canine Aspergillosis - OMICS Publishing Group

Canine Aspergillosis - OMICS Publishing Group

BROADLY REACTIVE PAN-VIRAL PCR OF CEREBROSPINAL FLUID IN CANINE MENINGOENCEPHALITIS OF UNKNOWN ETIOLOGY The canine meningoencephalitides of unknown etiology (MUE) GME,NME,NLE Histopathologic lesions are similar to those present in human viral meningoencephalitis PCR method has demonstrated that 50-70% of human meningoencephalitides are caused by CNS viral infections. We hypothesize that a subset of canine MUE results from aberrant immune responses following infection of the CNS. Objective : Objective : To determine whether or not nucleic acids from infectious agents can be identified in cerebrospinal fluid (CSF) by applying degenerate viral PCR to 146 CSF samples, collected pre- and/or postmortem from dogs with MUE and control dog. Consensus Degenerate Primers (CODEHOP)


C C T T A T C G C TG T 5 Consensus Clamp 3 Degenerate Core

Rose et al., 1998, Nucl Acids Res 26:1628 Viral Families: Herpesviridae, Adenoviridae, Alphaviridae, Picornaviridae, Paramyxoviridae, Polyomaviridae, Flaviviridae, Bunyaviridae, Bornaviridae, Rhabdoviridae, Coronaviridae Pan-Viral PCR Positive Controls Methods DNA and RNA extracted from 146 CSF samples and non-neurological controls by standard methods (Qiagen and Invitrogen) Housekeeping PCR and RT PCR for GAPDH (DNA) and beta-actin to confirm DNA and RNA integrity PCR and RT PCR on ~ 5 l of each sample in 20 various CODEHOP reactions Broadly reactive pan-viral PCR on MUE CSF (146 cases) Coronavirus

Bornavirus Rhabdovirus Alphavirus Herpesvirus Flavivirus Pan-bunyavirus PCR LaCrosse virus Breed CSF from 6/60 (10%) MUE cases positive on panbunyavirus PCR Sequences analysis disclosed 99% homology to LAC Black et al. J Vet Diagn Invest. 1994 Apr;6(2):250-4. Specific LAC PCR underway Developing an Ab to

LaCrosse to perform serology on CSF and serum in dog Shih Tzu Sex Color Weight M/N Wh/ Tan 7.4kg CSF RBC=1963 WBC=12 TP=23.8 Mixed pleocytosis MRI Diagnosis MUE (brain)

normal brain Pug F/S Black 8.25kg Weimaraner F/S Gray 25.8kg RBC=87 WBC=9 TP=17.0 Macro56%,Neut1 8%, Lymph16%, Eos10%

RBC=0 WBC=3 TP=38.7 RBC=38 WBC=26 TP=96.1 Lymph85%, Macro13%, neut2% 9.3kg RBC 69 WBC 20 TP 16.5; 2%% nondeg neut 19% lg mono 79% sm lymphs normal brain MUE (brain) 3.8kg WBC 1 TP

19.5 ;1% nondeg neut 26% lg mono 73% sm lymphs T2W multifocal hyperintense lesions throughout brain MUE (brain) Boxer Boston Terrier Chihuahua M M F/S

Fawn Blk/Wh Wh 37kg T2W hyperintensity cingulate gyrus T2W IM hyperintensity thoracic cord N/D Bunyavirus brain tumor +/meningitis MUE (T/L) MUE (brain) Pan-polyomavirus PCR Merkel Cell

Polyomavirus CSF from 3/60 (5%) MUE cases positive on panpolyomavirus PCR Sequencing (320 bp) shows 98% homology to Merkel Cell Polyomavirus (MCV) Feng et al. Science. 2008 Feb 22;319(5866): 1096-100. IHC on one case negative with human MCV Ab Specific MCV PCR underway CSF antigen testing Labrador Retriever Border Collie Great Dane F/I Chocol ate

26 kg M/N Blk/Wht 18.2 kg M/N RBC=1263 WBC=11 TP=19 16% Lymphs 10%Monos 73%Neuts 1%Eos RBC = 30, WBC = 139 TP 28 mixed pleocytosis RBC 4 WBC 53 TP 56 17% monos 52% eos 31% lymphs Normal cervical spine.

MUE (cervical) Hydrocephalus and syringohydromyelia MUE Syringohydromyelia MUE

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