Improving newborn screening for cystic fibrosis using next-generation sequencing technology: a technical feasibility...

Area of Science:

• Medical Genetics
• Newborn Screening
• Molecular Diagnostics

Background:

• Newborn screening (NBS) for cystic fibrosis (CF) often uses immunoreactive trypsinogen (IRT) with limited cystic fibrosis transmembrane regulator (CFTR) mutation panels.
• Improving the sensitivity and specificity of CF NBS is crucial for early diagnosis and intervention.

Purpose of the Study:

• To evaluate the feasibility and performance of a next-generation sequencing (NGS) assay for detecting CFTR mutations in newborn screening.
• To assess the potential of an IRT/NGS screening algorithm for improved CF diagnosis.

Main Methods:

• Developed and validated an NGS assay to detect 162 CFTR mutations/variants using DNA from dried blood spots (DBSs).
• Retrospectively analyzed 165 CF screen-positive infant samples with one known CFTR mutation.

Main Results:

• The NGS assay demonstrated 100% concordance with traditional methods, accurately detecting all validated CFTR mutations from DBS DNA.
• Identified nine infants with two CF-causing mutations and five with mutations outside the initial panel, improving diagnostic yield.
• The IRT/NGS algorithm showed potential for enhanced sensitivity, specificity, and positive predictive value (PPV).

Conclusions:

• The validated NGS assay is a reliable tool for CFTR mutation detection in NBS.
• Implementing an IRT/NGS strategy can significantly improve the accuracy and effectiveness of newborn screening for cystic fibrosis.
• This study provides a foundation for adopting NGS in routine NBS laboratories.