Faecal calprotectin and gut microbiota do not predict enteropathy in very preterm infants

Abstract

Aim: Very preterm birth is associated with a high risk of enteropathies. Diagnosis is challenging, especially in mild forms, leading to unnecessary periods of cessation of enteral feeding. This study aimed at establishing a prognosis score of enteropathy combining clinical parameters and faecal calprotectin concentration.

Methods: This prospective multicentric study included preterm neonates born at a gestational age of 33 weeks or less. Stools were collected weekly until hospital discharge, and daily in case of digestive events for calprotectin measurement (ELISA and immunochromatography) and microbiota analyses (16S rRNA gene sequencing).

Results: Among the 121 neonates included, 21 experienced at least one episode of enteropathy, mainly mild forms. By ELISA testing, median faecal calprotectin was 88 (8-798) µg/g faeces. No statistically significant association was found between the outset of enteropathy and maternal and neonatal characteristics, and calprotectin levels. The agreement between ELISA and immunochromatography assay was moderate (intra-class correlation coefficient 0.58, 95%CI [0.47-0.66]). Comparison of species diversity and relative bacterial abundance profiles between infants with or without enteropathy revealed no specific alterations associated with enteropathy.

Conclusion: The study failed to propose a prognostic score of enteropathy, probably due the large inter- and intra-individual variability of faecal calprotectin in very preterm neonates.

Trial registration: ClinicalTrials.gov NCT02010268.

Keywords: enteropathy; faecal calprotectin; gut microbiota; necrotising enterocolitis; preterm neonates.

FIGURE 1

Individual evolution of faecal calprotectin in the 21 neonates suffering from enteropathy event(s). Events are identified by the first day (D) of enteral feeding discontinuation (vertical dotted line): 18 patients with mild enteropathy (suspected NEC, Bell stage Ia and Bell stage Ib), and 5 with severe enteropathy (definite NEC, patients 80 (D71), 86, 120 and 121: Bell stage IIA; patient 104: Bell stage IIb). Blue line: faecal calprotectin (FC) by ELISA; pink line: FC by immunochromatographic (IC) assay

FIGURE 2

Microbiota profiles in neonates suffering from enteropathy and matching controls. A, Relative abundance of sequence reads at the phylum level assigned to different bacterial taxa in each neonate. B, Bacterial alpha‐diversity using the Chao‐1, Shannon and Inverse Simpson Diversity Indexes in control and neonates suffering from enteropathy. C, Beta‐diversity was analysed using multidimensional scaling (MDS) plot of samples according to disease status (enteropathy vs control) on the weighted‐UniFrac distance metrics