Accuracy of Immunofluorescence in the Diagnosis of Primary Ciliary Dyskinesia

Amelia Shoemark (Lead / Corresponding author), Emily Frost, Mellisa Dixon, Sarah Ollosson, Kate Kilpin, Mitali Patel, Juliet Scully, Andrew V. Rogers, Hannah M. Mitchison, Andrew Bush, Claire Hogg

Research output: Contribution to journalArticle

30 Citations (Scopus)
65 Downloads (Pure)

Abstract

Rationale The standard approach to diagnosis of primary ciliary dyskinesia (PCD) in the UK consists of assessing ciliary function by high-speed-microscopy and ultrastructure by election microscopy, but equipment and expertise is not widely available internationally. The identification of bi-allelic disease causing mutations is also diagnostic, but many disease causing genes are unknown, and testing is not widely available outside the USA. Fluorescent antibodies to ciliary proteins are used to validate research genetic studies, but diagnostic utility in this disease has not been systematically evaluated. Objectives Determine utility of a panel of six fluorescent labelled antibodies as a diagnostic tool for PCD. Methods Immunofluorescent labelling of nasal brushings from a discovery cohort of 35 patients diagnosed with PCD by ciliary ultrastructure, and a diagnostic accuracy cohort of 386 patients referred with symptoms suggestive of disease. The results were compared to diagnostic outcome. Measurements and Main Results Immunofluorescence correctly identified mislocalised or absent staining in 100% of the discovery cohort. In the diagnostic cohort immunofluorescence successfully identified 22 of 25 patients with PCD and normal staining in all 252 in whom PCD was considered highly unlikely. Immunofluorescence additionally provided a result in 55% (39) of cases which were previously inconclusive. Immunofluorescence results were available within 14 days, costing $187 per sample compared to electron microscopy (27 days, cost $1452). Conclusions Immunofluorescence is a highly specific diagnostic test for PCD, and improves the speed and availability of diagnostic testing, however, sensitivity is limited and immunofluorescence is not suitable as a stand-alone test.

Original languageEnglish
Pages (from-to)94-101
Number of pages8
JournalAmerican Journal of Respiratory and Critical Care Medicine
Volume196
Issue number1
Early online date15 Feb 2017
DOIs
Publication statusPublished - 1 Jul 2017

Fingerprint

Kartagener Syndrome
Fluorescent Antibody Technique
Microscopy
Staining and Labeling
Genetic Research
Antibodies
Nose
Routine Diagnostic Tests
Electron Microscopy
Costs and Cost Analysis
Equipment and Supplies
Mutation
Genes

Keywords

  • CILIA
  • Ultrastructure
  • ANTIBODY
  • respiratory epithelium

Cite this

Shoemark, Amelia ; Frost, Emily ; Dixon, Mellisa ; Ollosson, Sarah ; Kilpin, Kate ; Patel, Mitali ; Scully, Juliet ; Rogers, Andrew V. ; Mitchison, Hannah M. ; Bush, Andrew ; Hogg, Claire. / Accuracy of Immunofluorescence in the Diagnosis of Primary Ciliary Dyskinesia. In: American Journal of Respiratory and Critical Care Medicine. 2017 ; Vol. 196, No. 1. pp. 94-101.
@article{08333f902dc7433cab935c6d2954c2bb,
title = "Accuracy of Immunofluorescence in the Diagnosis of Primary Ciliary Dyskinesia",
abstract = "Rationale The standard approach to diagnosis of primary ciliary dyskinesia (PCD) in the UK consists of assessing ciliary function by high-speed-microscopy and ultrastructure by election microscopy, but equipment and expertise is not widely available internationally. The identification of bi-allelic disease causing mutations is also diagnostic, but many disease causing genes are unknown, and testing is not widely available outside the USA. Fluorescent antibodies to ciliary proteins are used to validate research genetic studies, but diagnostic utility in this disease has not been systematically evaluated. Objectives Determine utility of a panel of six fluorescent labelled antibodies as a diagnostic tool for PCD. Methods Immunofluorescent labelling of nasal brushings from a discovery cohort of 35 patients diagnosed with PCD by ciliary ultrastructure, and a diagnostic accuracy cohort of 386 patients referred with symptoms suggestive of disease. The results were compared to diagnostic outcome. Measurements and Main Results Immunofluorescence correctly identified mislocalised or absent staining in 100{\%} of the discovery cohort. In the diagnostic cohort immunofluorescence successfully identified 22 of 25 patients with PCD and normal staining in all 252 in whom PCD was considered highly unlikely. Immunofluorescence additionally provided a result in 55{\%} (39) of cases which were previously inconclusive. Immunofluorescence results were available within 14 days, costing $187 per sample compared to electron microscopy (27 days, cost $1452). Conclusions Immunofluorescence is a highly specific diagnostic test for PCD, and improves the speed and availability of diagnostic testing, however, sensitivity is limited and immunofluorescence is not suitable as a stand-alone test.",
keywords = "CILIA, Ultrastructure, ANTIBODY, respiratory epithelium",
author = "Amelia Shoemark and Emily Frost and Mellisa Dixon and Sarah Ollosson and Kate Kilpin and Mitali Patel and Juliet Scully and Rogers, {Andrew V.} and Mitchison, {Hannah M.} and Andrew Bush and Claire Hogg",
year = "2017",
month = "7",
day = "1",
doi = "10.1164/rccm.201607-1351OC",
language = "English",
volume = "196",
pages = "94--101",
journal = "American Journal of Respiratory and Critical Care Medicine",
issn = "1073-449X",
publisher = "American Thoracic Society",
number = "1",

}

Shoemark, A, Frost, E, Dixon, M, Ollosson, S, Kilpin, K, Patel, M, Scully, J, Rogers, AV, Mitchison, HM, Bush, A & Hogg, C 2017, 'Accuracy of Immunofluorescence in the Diagnosis of Primary Ciliary Dyskinesia', American Journal of Respiratory and Critical Care Medicine, vol. 196, no. 1, pp. 94-101. https://doi.org/10.1164/rccm.201607-1351OC

Accuracy of Immunofluorescence in the Diagnosis of Primary Ciliary Dyskinesia. / Shoemark, Amelia (Lead / Corresponding author); Frost, Emily; Dixon, Mellisa; Ollosson, Sarah; Kilpin, Kate; Patel, Mitali; Scully, Juliet; Rogers, Andrew V.; Mitchison, Hannah M.; Bush, Andrew; Hogg, Claire.

In: American Journal of Respiratory and Critical Care Medicine, Vol. 196, No. 1, 01.07.2017, p. 94-101.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Accuracy of Immunofluorescence in the Diagnosis of Primary Ciliary Dyskinesia

AU - Shoemark, Amelia

AU - Frost, Emily

AU - Dixon, Mellisa

AU - Ollosson, Sarah

AU - Kilpin, Kate

AU - Patel, Mitali

AU - Scully, Juliet

AU - Rogers, Andrew V.

AU - Mitchison, Hannah M.

AU - Bush, Andrew

AU - Hogg, Claire

PY - 2017/7/1

Y1 - 2017/7/1

N2 - Rationale The standard approach to diagnosis of primary ciliary dyskinesia (PCD) in the UK consists of assessing ciliary function by high-speed-microscopy and ultrastructure by election microscopy, but equipment and expertise is not widely available internationally. The identification of bi-allelic disease causing mutations is also diagnostic, but many disease causing genes are unknown, and testing is not widely available outside the USA. Fluorescent antibodies to ciliary proteins are used to validate research genetic studies, but diagnostic utility in this disease has not been systematically evaluated. Objectives Determine utility of a panel of six fluorescent labelled antibodies as a diagnostic tool for PCD. Methods Immunofluorescent labelling of nasal brushings from a discovery cohort of 35 patients diagnosed with PCD by ciliary ultrastructure, and a diagnostic accuracy cohort of 386 patients referred with symptoms suggestive of disease. The results were compared to diagnostic outcome. Measurements and Main Results Immunofluorescence correctly identified mislocalised or absent staining in 100% of the discovery cohort. In the diagnostic cohort immunofluorescence successfully identified 22 of 25 patients with PCD and normal staining in all 252 in whom PCD was considered highly unlikely. Immunofluorescence additionally provided a result in 55% (39) of cases which were previously inconclusive. Immunofluorescence results were available within 14 days, costing $187 per sample compared to electron microscopy (27 days, cost $1452). Conclusions Immunofluorescence is a highly specific diagnostic test for PCD, and improves the speed and availability of diagnostic testing, however, sensitivity is limited and immunofluorescence is not suitable as a stand-alone test.

AB - Rationale The standard approach to diagnosis of primary ciliary dyskinesia (PCD) in the UK consists of assessing ciliary function by high-speed-microscopy and ultrastructure by election microscopy, but equipment and expertise is not widely available internationally. The identification of bi-allelic disease causing mutations is also diagnostic, but many disease causing genes are unknown, and testing is not widely available outside the USA. Fluorescent antibodies to ciliary proteins are used to validate research genetic studies, but diagnostic utility in this disease has not been systematically evaluated. Objectives Determine utility of a panel of six fluorescent labelled antibodies as a diagnostic tool for PCD. Methods Immunofluorescent labelling of nasal brushings from a discovery cohort of 35 patients diagnosed with PCD by ciliary ultrastructure, and a diagnostic accuracy cohort of 386 patients referred with symptoms suggestive of disease. The results were compared to diagnostic outcome. Measurements and Main Results Immunofluorescence correctly identified mislocalised or absent staining in 100% of the discovery cohort. In the diagnostic cohort immunofluorescence successfully identified 22 of 25 patients with PCD and normal staining in all 252 in whom PCD was considered highly unlikely. Immunofluorescence additionally provided a result in 55% (39) of cases which were previously inconclusive. Immunofluorescence results were available within 14 days, costing $187 per sample compared to electron microscopy (27 days, cost $1452). Conclusions Immunofluorescence is a highly specific diagnostic test for PCD, and improves the speed and availability of diagnostic testing, however, sensitivity is limited and immunofluorescence is not suitable as a stand-alone test.

KW - CILIA

KW - Ultrastructure

KW - ANTIBODY

KW - respiratory epithelium

U2 - 10.1164/rccm.201607-1351OC

DO - 10.1164/rccm.201607-1351OC

M3 - Article

VL - 196

SP - 94

EP - 101

JO - American Journal of Respiratory and Critical Care Medicine

JF - American Journal of Respiratory and Critical Care Medicine

SN - 1073-449X

IS - 1

ER -