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Background  No definitive, antemortem diagnostic test for canine degenerative myelopathy ( DM ) is available. Phosphorylated neurofilament heavy ( pNF ‐H) is a promising biomarker for nervous system diseases.    Hypothesis/Objective  Cerebrospinal fluid ( CSF ) and serum  pNF ‐H is a detectable biological marker for diagnosis of canine  DM .    Animals  Fifty‐three  DM ‐affected, 27 neurologically normal, 7 asymptomatic at‐risk, and 12  DM  mimic dogs.    Methods  Archived  CSF  and serum  pNF ‐H concentrations were determined by a commercially available  ELISA . A receiver‐operating characteristic ( ROC ) curve was generated with  CSF  values.    Results  Compared with old control dogs, median  CSF pNF ‐H concentration was increased in all stages of  DM ; old dogs 5.1 ng/mL (interquartile range [ IQR ] 1.4–9.3) versus  DM  stage 1 23.9 ng/mL ( IQR  20.8–29.6;  P  < .05) versus  DM  stage 2 36.8 ng/mL ( IQR  22.9–51.2;  P  < .0001) versus  DM  stage 3 25.2 ng/mL ( IQR  20.2–61.8;  P  < .001) versus  DM  stage 4 38.0 ng/mL ( IQR  11.6–59.9;  P  < .01). Degenerative myelopathy stage 1 dogs had increased median  CSF pNF ‐H concentrations compared with asymptomatic, at‐risk dogs (3.4 ng/mL [ IQR  1.5–10.9;  P  < .01]) and  DM  mimics (6.6 ng/mL [ IQR  3.0–12.3;  P  < .01]).  CSF pNF ‐H concentration >20.25 ng/mL was 80.4% sensitive (confidence interval [ CI ] 66.09–90.64%) and 93.6% specific ( CI  78.58–99.21%) for  DM . Area under the  ROC  curve was 0.9467 ( CI  0.92–0.9974). No differences in serum  pNF ‐H concentration were found between control and  DM ‐affected dogs.    Conclusions and Clinical Importance   pNF ‐H concentration in  CSF  is a sensitive biomarker for diagnosis of  DM . Although there was high specificity for  DM  in this cohort, further study should focus on a larger cohort of  DM  mimics, particularly other central and peripheral axonopathies.

Cerebrospinal Fluid Levels of Phosphorylated Neurofilament Heavy as a Diagnostic Marker of Canine Degenerative Myelopathy