Utility of cell-free DNA concentrations and illness severity scores to predict survival in critically ill neonatal foals

Abstract

Plasma cell-free DNA (cfDNA) levels have been associated with disease and survival status in septic humans and dogs. To date, studies investigating cfDNA levels in association with critical illness in foals are lacking. We hypothesized that cfDNA would be detectable in the plasma of foals, that septic and sick-nonseptic foals would have significantly higher cfDNA levels compared to healthy foals, and that increased cfDNA levels would be associated with non-survival. Animals used include 80 foals of 10 days of age or less admitted to a tertiary referral center between January and July, 2020 were stratified into three categories: healthy (n = 34), sick non-septic (n = 11) and septic (n = 35) based on specific criteria. This was a prospective clinical study. Blood was collected from critically ill foals at admission or born in hospital for cfDNA quantification and blood culture. Previously published sepsis score (SS) and neonatal SIRS score (NSIRS) were also calculated. SS, NSIRS, blood culture status and cfDNA concentrations were evaluated to predict survival. Continuous variables between groups were compared using Kruskal-Wallis ANOVA with Dunn's post hoc test. Comparisons between two groups were assessed using the Mann-Whitney U-test or Spearman rank for correlations. The performance of cfDNA, sepsis score and NSIRS score to predict survival was assessed by receiver operator characteristic (ROC) curve analysis including area under the curve, sensitivity and specificity using cutoffs. Plasma cfDNA was detectable in all foals. No significant differences in cfDNA concentration were detected between healthy foals and septic foals (P = 0.65) or healthy foals and sick non-septic foals (P = 0.88). There was no significant association between cfDNA and culture status, SS, NSIRS or foal survival. SS (AUC 0.85) and NSIRS (AUC 0.83) were superior to cfDNA (AUC 0.64) in predicting survival. Although cfDNA was detectable in foal plasma, it offers negligible utility to diagnose sepsis or predict survival in critical illness in neonates.

Fig 1. Plasma cell-free DNA (cfDNA) concentrations in healthy foals (n = 34), sick non-septic foals (n = 11) and septic foals (n = 35) based on defined criteria (sepsis score and/or blood culture status).

Symbols represent values for individual foals with median and interquartile range error bars (P>0.05 between groups).

Fig 2. Plasma cell-free DNA (cfDNA) concentrations and illness severity scores in 80 hospitalized foals.

2A: cfDNA concentration and sepsis score (r = 0.07; P = 0.48). 2B: cfDNA concentration and NSIRS score (r = 0.07; P = 0.54). Symbols represent values for individual foals.

Fig 3. Plasma cell-free DNA (cfDNA) concentrations in 80 hospitalized neonatal foals with positive blood culture (n = 24) and negative blood culture (n = 56).

Symbols represent values for individual foals with median and interquartile range error bars (P = 0.07).

Fig 4. Plasma cell-free DNA (cfDNA) concentrations in 80 hospitalized neonatal foals that survived (n = 69) and died (n = 11).

Symbols represent values for individual foals with median and interquartile range error bars (P = 0.16).

Fig 5

Receiver operator characteristic (ROC) curves for cfDNA (A), sepsis score (B) and NSIRS score (C) for predicting survival in 80 hospitalized neonatal foals.