1d011292198ddf645766fcfb0cc6354f pmc_33182294.pdf ff2f426292b264275307d3809d08a43f3fc8efdd pmc_33182294.pdf 6aac101c59a705d3efefbee1dc47c080f15f0828abccecf96898e7018e28d546 pmc_33182294.pdf Title: Immunofluorescence Analysis as a Diagnostic Tool in a Spanish Cohort of Patients with Suspected Primary Ciliary Dyskinesia Subject: 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. Keywords: cilia; primary ciliary dyskinesia; PCD; immunofluorescence; antibody Author: Noelia Baz-Redón, Sandra Rovira-Amigo, Mónica Fernández-Cancio, Silvia Castillo-Corullón, Maria Cols, M. Araceli Caballero-Rabasco, Óscar Asensio, Carlos Martín de Vicente, Maria del Mar Martínez-Colls, Alba Torrent-Vernetta, Inés de Mir-Messa, Silvia Gartner, Ignacio Iglesias-Serrano, Ana Díez-Izquierdo, Eva Polverino, Esther Amengual-Pieras, Rosanel Amaro-Rodríguez, Montserrat Vendrell, Marta Mumany, María Teresa Pascual-Sánchez, Belén Pérez-Dueñas, Ana Reula, Amparo Escribano, Francisco Dasí, Miguel Armengot-Carceller, Marta Garrido-Pontnou, Núria Camats-Tarruella and Antonio Moreno-Galdó Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Wed Nov 18 13:25:02 2020 CET ModDate: Wed Nov 18 13:28:06 2020 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 7306180 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- GFKJOY+VnURWPalladioL Type 1 Custom yes yes no 10 0 IRAVWY+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 DGQEMW+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 HZRVFI+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 FATHTC+Rpxr Type 1 Builtin yes yes no 28 0 MMXMSZ+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 CUAJJQ+Pxbsy Type 1 Builtin yes yes no 36 0 XPUSBJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 44 0 BSDWLC+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 47 0 IUBRKP+Pxsy Type 1 Builtin yes yes no 69 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 76 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 80 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 84 0 OMJGHE+PalatinoLinotype TrueType WinAnsi yes yes no 220 0 OMJGHG+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 223 0 OMJGII+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 226 0 OMJGNI+TimesNewRoman CID TrueType Identity-H yes yes yes 229 0 TGMEOR+Rpcxr Type 1 Builtin yes yes no 243 0 RQODNN+Rpxb Type 1 Builtin yes yes no 250 0 URYLRZ+URWPalladioL-BoldItal Type 1 Custom yes yes no 254 0 CKBPVA+EURM10 Type 1 Builtin yes yes no 261 0 OMJGHD+PalatinoLinotype CID TrueType Identity-H yes yes yes 26646 0 OMJGHE+PalatinoLinotype TrueType WinAnsi yes yes no 26652 0 OMJGHG+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 26655 0 OMJGII+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 26658 0 OMJGNI+TimesNewRoman CID TrueType Identity-H yes yes yes 26661 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-02 04:17:48 CEST RepresentationInformation: pmc_33182294.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 11:43:32 CEST Size: 7306180 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 26898 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Experimental Section Destination: section.2 Children: Item: Title: Patients Destination: subsection.2.1 Item: Title: PCD Diagnostic Evaluation Destination: subsection.2.2 Item: Title: Immunofluorescence Technique and Analysis Destination: subsection.2.3 Item: Title: Data Analysis Destination: subsection.2.4 Item: Title: Results Destination: section.3 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: section.4 Info: Title: Immunofluorescence Analysis as a Diagnostic Tool in a Spanish Cohort of Patients with Suspected Primary Ciliary Dyskinesia Author: Noelia Baz-Redón, Sandra Rovira-Amigo, Mónica Fernández-Cancio, Silvia Castillo-Corullón, Maria Cols, M. Araceli Caballero-Rabasco, Óscar Asensio, Carlos Martín de Vicente, Maria del Mar Martínez-Colls, Alba Torrent-Vernetta, Inés de Mir-Messa, Silvia Gartner, Ignacio Iglesias-Serrano, Ana Díez-Izquierdo, Eva Polverino, Esther Amengual-Pieras, Rosanel Amaro-Rodríguez, Montserrat Vendrell, Marta Mumany, María Teresa Pascual-Sánchez, Belén Pérez-Dueñas, Ana Reula, Amparo Escribano, Francisco Dasí, Miguel Armengot-Carceller, Marta Garrido-Pontnou, Núria Camats-Tarruella and Antonio Moreno-Galdó Subject: 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. 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