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None Fits All: Unraveling Structural Rearrangements of the CFH Gene Cluster in Atypical Hemolytic Uremic Syndrome Patients Using Molecular Combing and Long-Fragment Targeted Sequencing

Lookup NU author(s): Dr Patrick Walsh, Professor David KavanaghORCiD



This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND).


Complement factor H (CFH) and its related proteins have an essential role in regulating the alternative pathway of the complement system. Mutations and structural variants (SVs) of the CFH gene cluster, consisting of CFH and its five related genes (CFHR1-5), have been reported in renal pathologies as well as in complex immune diseases like age-related macular degeneration and systemic lupus erythematosus. SV analysis of this cluster is challenging because of its high degree of sequence homology. Following first-line next-generation sequencing gene panel sequencing, we applied Genomic Vision's Molecular Combing Technology to detect and visualize SVs within the CFH gene cluster and resolve its structural haplotypes completely. This approach was tested in three patients with atypical hemolytic uremic syndrome and known SVs and 18 patients with atypical hemolytic uremic syndrome or complement factor 3 glomerulopathy with unknown CFH gene cluster haplotypes. Three SVs, a CFH/CFHR1 hybrid gene in two patients and a rare heterozygous CFHR4/CFHR1 deletion in trans with the common CFHR3/CFHR1 deletion in a third patient, were newly identified. For the latter, the breakpoints were determined using a targeted enrichment approach for long DNA fragments (Samplix Xdrop) in combination with Oxford Nanopore sequencing. Molecular combing in addition to next-generation sequencing was able to improve the molecular genetic yield in this pilot study. This (cost-)effective approach warrants validation in larger cohorts with CFH/CFHR-associated disease.

Publication metadata

Author(s): Tschernoster N, Erger F, Walsh P, Mhaille B, Kavanagh D, Fistrek M, Habbig S, Kukat C, Schumacher L, Folz K, Tolia MR, Becker C, Thiele H, Nuernberg P, Beck B, Altmüller J

Publication type: Article

Publication status: Published

Journal: The Journal of Molecular Diagnostics

Year: 2022

Volume: 24

Issue: 6

Pages: 619-631

Print publication date: 01/06/2022

Online publication date: 07/04/2022

Acceptance date: 25/02/2022

Date deposited: 03/07/2023

ISSN (electronic): 1525-1578

Publisher: Elsevier


DOI: 10.1016/j.jmoldx.2022.02.006

PubMed id: 35398599


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