Evaluation of Open-Source Ciliary Analysis Software in Primary Ciliary Dyskinesia: A Comparative Assessment

Primary Ciliary Dyskinesia (PCD) is a rare genetic disorder characterized by alterations in motile cilia function. The diagnosis of PCD is challenging due to the lack of standardized methods in clinical practice. High-speed video microscopy analysis (HSVA) directly evaluates ciliary beat frequency (CBF) in PCD. Recently, open-source ciliary analysis software applications have shown promise in measuring CBF accurately. However, there is limited knowledge about the performance of different software applications, creating a gap in understanding their comparative effectiveness in measuring CBF in PCD. We compared two open-source software applications, CiliarMove (v219) and Cilialyzer (v1.2.1-b3098cb), against the manual count method. We used high-speed videos of nasal ciliary brush samples from PCD <i>RSPH4A</i>-positive (PCD (<i>RSPH4A</i>)) patients and healthy controls. All three methods showed lower median CBF values for patients with PCD (<i>RSPH4A</i>) than in healthy controls. CiliarMove and Cilialyzer identified lower CBF in patients with PCD (<i>RSPH4A</i>), similarly to the manual count. Cilialyzer, CiliarMove, and manual count methods demonstrated statistical significance (<i>p</i>-value < 0.0001) in the difference of median CBF values between patients with PCD (<i>RSPH4A</i>) and healthy controls. Correlation coefficients between the manual count values against both software methods demonstrated positive linear relationships. These findings support the utility of open-source software-based analysis tools. Further studies are needed to validate these findings with other genetic variants and identify the optimal software for accurate CBF measurement in patients with PCD.