Reclassifying inconclusive diagnosis after newborn screening for cystic fibrosis. Moving forward

Published:December 21, 2021DOI:


      • This study investigates the association of cystic fibrosis or CFTR-Related disorders (CFTR-RD) in babies with inconclusive diagnosis at newborn screening.
      • We performed whole CFTR gene sequencing, in vivo CFTR function measurement, and in vitro studies of variants of unknown significance.
      • This workflow differentiated children at high likelihood of evolving to CF, those at extremely low risk of developing a CFTR-RD and those with partial CFTR dysfunction who might develop CFTR-RD later on.



      Newborn screening for Cystic Fibrosis (CF) is associated with situations where the diagnosis of CF or CFTR related disorders (CFTR-RD) cannot be clearly ruled out.

      Materials/patients and methods

      We report a case series of 23 children with unconclusive diagnosis after newborn screening for CF and a mean follow-up of 7.7 years (4–13). Comprehensive investigations including whole CFTR gene sequencing, in vivo intestinal current measurement (ICM), nasal potential difference (NPD), and in vitro functional studies of variants of unknown significance, helped to reclassify the patients.


      Extensive genetic testing identified, in trans with a CF causing mutation, variants with varying clinical consequences and 3 variants of unknown significance (VUS). Eighteen deep intronic variants were identified by deep resequencing of the whole CFTR gene in 13 patients and were finally considered as non-pathogenic. All patients had normal CFTR dependent chloride transport in ICM. NPD differentiated 3 different profiles: CF-like tracings qualifying the patients as CF, such as F508del/D1152H patients; normal responses, suggesting an extremely low likelihood of developing a CFTR-RD such as F508del/TG11T5 patients; partial CFTR dysfunction above 20% of the normal, highlighting a remaining risk of developing CFTR-RD such as F508del/F1052V patients. The 3 VUS were reclassified as variant with defective maturation (D537N), defective expression (T582I) or with no clinical consequence (M952T).


      This study demonstrates the usefulness of combining genetic and functional investigations to assess the possibility of evolving to CF or CFTR-RD in babies with inconclusive diagnosis at neonatal screening.


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