Microarray-based mutation analysis of 183 Spanish families with Usher syndrome.
OBJECTIVE
The purpose of this study was to test the ability of the genotyping microarray for Usher syndrome (USH) to identify the mutations responsible for the disease in a cohort of 183 patients with USH.


METHODS
DNA from 183 patients with Usher syndrome from the Spanish population was analyzed using a genotyping microarray containing 429 previously identified disease-associated variants in eight USH genes. Mutations detected by the array were confirmed by direct sequencing. Haplotype analysis was also performed in families carrying common Spanish mutations.


RESULTS
The genotyping microarray identified 43 different variants, divided into 32 disease causative and 11 probably nonpathologic. Mutations were detected in 62 patients with USH (33.9%). According to the clinical classification of patients, pathologic variants were detected in 31.4% patients with USH1, 39.4% of with USH2, 22.2% with USH3 and 15.8% with unclassified Usher syndrome. Ninety-seven pathologic alleles were detected, corresponding to 26.5% of expected alleles. The USH2A mutations p.C3267R and p.T3571M were revealed as common in the Spanish population, and two major haplotypes linked to these mutations were observed.


CONCLUSIONS
The genotyping microarray is a robust, low-cost, rapid technique that is effective for the genetic study of patients with USH. However, it also indicates variants of unclear pathologic nature and detection failures have also been observed. Results must be confirmed by direct sequencing to avoid misdiagnosis, and continuous updates of the microarray should be performed to increase the efficiency and rate of detection of mutations.
Jaijo
Teresa
T
Unidad de Genética. Hospital Universitario La Fe, Valencia, Spain.
Aller
Elena
E
García-García
Gema
G
Aparisi
María J
MJ
Bernal
Sara
S
Avila-Fernández
Almudena
A
Barragán
Isabel
I
Baiget
Montserrat
M
Ayuso
Carmen
C
Antiñolo
Guillermo
G
Díaz-Llopis
Manuel
M
Külm
Maigi
M
Beneyto
Magdalena
M
Nájera
Carmen
C
Millán
Jose M
JM
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D000483:Alleles; D000070585:Cadherin Related Proteins; D015820:Cadherins / Q000235:genetics; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D004252:DNA Mutational Analysis; D016326:Extracellular Matrix Proteins / Q000235:genetics; D020869:Gene Expression Profiling; D005838:Genotype; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D009154:Mutation; D000081412:Myosin VIIa; D009218:Myosins / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D043562:Receptors, G-Protein-Coupled / Q000235:genetics; D013030:Spain; D052245:Usher Syndromes / Q000235:genetics*
C417525:ADGRV1 protein, human; D048868:Adaptor Proteins, Signal Transducing; C442480:CDH23 protein, human; C498835:CDHR15, human; C437067:CLRN1 protein, human; D000070585:Cadherin Related Proteins; D015820:Cadherins; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D016326:Extracellular Matrix Proteins; C000631455:MYO7A protein, human; D008565:Membrane Proteins; D000081412:Myosin VIIa; D009419:Nerve Tissue Proteins; D043562:Receptors, G-Protein-Coupled; C118970:USH1C protein, human; C477452:USH1G protein, human; C112735:USH2A protein, human; D009218:Myosins