| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1From the Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan; 2McGill Ocular Genetics Laboratory, McGill University Health Center, MTL Children’s Hospital, Montreal, Quebec, Canada; and the 3National Eye Institute, National Institutes of Health, Bethesda, Maryland.
PURPOSE. To develop and apply microarray-based resequencing technology to detect sequence alterations in multiple autosomal recessive retinal disease genes on a single high-throughput platform.
METHODS. Oligonucleotides corresponding to both strands of the target exons and the flanking intron sequences of 29,214 bp from 11 genes associated with autosomal recessive retinitis pigmentosa (arRP) were tiled on 20 x 25-µm microarrays (arRP-I arrays). A total of 155 exons were amplified from 35 arRP patient DNA samples, with each sample being sequenced on an arRP-I chip by hybridization.
RESULTS. With the arRP-I arrays, 97.6% of the tiled sequence were determined with more than 99% accuracy and reproducibility. Of the 2.4% unread sequence, 89.5% involved stretches of G or C. In analyzing the 903,140-bp sequence from the 35 patient samples, 506 sequence changes have been detected in which 386 are previously reported alterations, and 120 are novel. In addition to four known causative mutations, six novel sequence changes that are potentially pathogenic were observed. Additional analysis is needed to determine whether these changes are responsible for arRP in these patients.
CONCLUSIONS. The use of microarray for sequencing is a novel approach, and the arRP-I chip is the first successful application of this technology for determining sequence alteration in multiple disease-related genes. These arrays can be used for high-throughput genotyping of patients with relevant retinal conditions. In addition, these arrays offer a unique opportunity to interrogate complex patterns of inheritance due to the involvement of more than one gene by screening multiple genes on a single platform.
This article has been cited by other articles:
|
|
 |
|
  A. I. den Hollander, J. J. C. van Lith-Verhoeven, M. L. Arends, T. M. Strom, F. P. M. Cremers, and C. B. Hoyng Novel Compound Heterozygous TULP1 Mutations in a Family With Severe Early-Onset Retinitis Pigmentosa Arch Ophthalmol, July 1, 2007; 125(7): 932 - 935. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. M. Riley, M. A. Mansilla, J. Ma, S. Daack-Hirsch, B. S. Maher, L. M. Raffensperger, E. T. Russo, A. R. Vieira, C. Dode, M. Mohammadi, et al. Impaired FGF signaling contributes to cleft lip and palate PNAS, March 13, 2007; 104(11): 4512 - 4517. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Daiger, S. J. Bowne, and L. S. Sullivan Perspective on Genes and Mutations Causing Retinitis Pigmentosa Arch Ophthalmol, February 1, 2007; 125(2): 151 - 158. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Blain and B. P. Brooks Molecular Diagnosis and Genetic Counseling in Ophthalmology Arch Ophthalmol, February 1, 2007; 125(2): 196 - 203. [Full Text] [PDF]
|
|
|
|
|
|
K. Downs, D. N. Zacks, R. Caruso, A. J. Karoukis, K. Branham, B. M. Yashar, M. H. Haimann, K. Trzupek, M. Meltzer, D. Blain, et al. Molecular Testing for Hereditary Retinal Disease as Part of Clinical Care Arch Ophthalmol, February 1, 2007; 125(2): 252 - 258. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
