Immunofluorescence Analysis as a Diagnostic Tool in a Spanish Cohort of Patients with Suspected Primary Ciliary Dyskinesia.

Noelia Baz-Redón,Marta Garrido-Pontnou,Ana Reula,Amparo Escribano,Núria Camats-Tarruella,Ana Díez-Izquierdo,Miguel Armengot-Carceller,Inés De Mir-Messa,Marta Mumany,Esther Amengual-Pieras,María Teresa Pascual-Sánchez,Eva Polverino,Montserrat Vendrell,Rosanel Amaro-Rodríguez,Carlos Martín De Vicente,Alba Torrent‐Vernetta ,Òscar Asensio ,María Cols ,Sílvia Gartner ,Belén Pérez‐Dueñas ,Sandra Rovira-Amigó ,Silvia Castillo‐Corullón ,María Del Mar Martínez-Colls ,Mónica Fernández‐Cancio ,M Araceli Caballero-Rabasco ,Ignacio Iglesias Serrano ,Francisco Dası́ ,Antonio Moreno‐Galdó

doi:10.3390/jcm9113603

Abstract

Primary ciliary dyskinesia (PCD) is an autosomal recessive rare disease caused by an alteration of ciliary structure. Immunofluorescence, consisting in the detection of the presence and distribution of cilia proteins in human respiratory cells by fluorescence, has been recently proposed as a technique to improve understanding of disease-causing genes and diagnosis rate in PCD. The objective of this study is to determine the accuracy of a panel of four fluorescently labeled antibodies (DNAH5, DNALI1, GAS8 and RSPH4A or RSPH9) as a PCD diagnostic tool in the absence of transmission electron microscopy analysis. The panel was tested in nasal brushing samples of 74 patients with clinical suspicion of PCD. Sixty-eight (91.9%) patients were evaluable for all tested antibodies. Thirty-three cases (44.6%) presented an absence or mislocation of protein in the ciliary axoneme (15 absent and 3 proximal distribution of DNAH5 in the ciliary axoneme, 3 absent DNAH5 and DNALI1, 7 absent DNALI1 and cytoplasmatic localization of GAS8, 1 absent GAS8, 3 absent RSPH9 and 1 absent RSPH4A). Fifteen patients had confirmed or highly likely PCD but normal immunofluorescence results (68.8% sensitivity and 100% specificity). In conclusion, immunofluorescence analysis is a quick, available, low-cost and reliable diagnostic test for PCD, although it cannot be used as a standalone test.

Highlights

Primary ciliary dyskinesia (PCD) is an autosomal recessive rare disease (1/15,000) caused by an alteration of ciliary structure, which impairs mucociliary clearance [1,2]
Immunofluorescence analysis was performed in a cohort of 74 PCD-suspected patients
After PCD diagnostic evaluation, 25 patients were considered as confirmed PCD, 25 as highly likely and 24 as highly unlikely PCD (Table 1)

Summary

Introduction

Primary ciliary dyskinesia (PCD) is an autosomal recessive rare disease (1/15,000) caused by an alteration of ciliary structure, which impairs mucociliary clearance [1,2]. As there is not a unique gold standard diagnostic test, the ERS [6] and the American Thoracic Society (ATS) [7] have recently proposed the use of different diagnostic techniques to improve the accuracy and diagnosis rate of PCD. Immunofluorescence (IF) is a technique consisting in the use of fluorescently labeled antibodies for the detection of the presence and distribution of different ciliary proteins in human respiratory cells by fluorescence or confocal microscopy, and it has been recently proposed as a tool to improve the diagnosis rate in PCD and facilitate a better understanding of disease-causing genes [6,7]. An important number of protein complexes are distributed among these microtubule structures: the outer (ODA) and inner dynein arms (IDA), the nexin links, the central sheath and the radial spokes [8,9] (Figure 1)

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Conclusion