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PURPOSE. Congenital stationary night blindness (CSNB) is a clinically and genetically heterogeneous retinal disease. Although electroretinographic (ERG) measurements can discriminate clinical subgroups, the identification of the underlying genetic defects has been complicated for CSNB because of genetic heterogeneity, the uncertainty about the mode of inheritance, and time-consuming and costly mutation scanning and direct sequencing approaches. METHODS. To overcome these challenges and to generate a time-and cost-efficient mutation screening tool, the authors developed a CSNB genotyping microarray with arrayed primer extension (APEX) technology. To cover as many mutations as possible, a comprehensive literature search was performed, and DNA samples from a cohort of patients with CSNB were first sequenced directly in known CSNB genes. Subsequently, oligonucleotides were designed representing 126 sequence variations in RHO, CABP4, CACNA1F, CACNA2D4, GNAT1, GRM6, NYX, PDE6B, and SAG and spotted on the chip. RESULTS. Direct sequencing of genes known to be associated with CSNB in the study cohort revealed 21 mutations (12 novel and 9 previously reported). The resultant microarray containing oligonucleotides, which allow to detect 126 known and novel mutations, was 100% effective in determining the expected sequence changes in all known samples assessed. In addition, investigation of 34 patients with CSNB who were previously not genotyped revealed sequence variants in 18%, of which 15% are thought to be disease-causing mutations. CONCLUSIONS. This relatively inexpensive first-pass genetic testing device for patients with a diagnosis of CSNB will improve molecular diagnostics and genetic counseling of patients and their families and gives the opportunity to analyze whether, for example, more progressive disorders such as cone or cone-rod dystrophies underlie the same gene defects. (Invest Ophthalmol Vis Sci. 2009;50:5919-5926) DOI:10.1167/iovs.09-3548
Keywords
ELECTRORETINOGRAM, RICH, COMPLETE FORM, FAMILIES, CACNA1F MUTATIONS, MUTATION DETECTION, STATIONARY NIGHT BLINDNESS, ARRAYED PRIMER EXTENSION, EXPRESSION, DYSTROPHY

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Chicago
Zeitz, Christina, Stephan Labs, Birgit Lorenz, Ursula Forster, Janne Uksti, Hester Y Kroes, Elfride De Baere, et al. 2009. “Genotyping Microarray for CSNB-associated Genes.” Investigative Ophthalmology & Visual Science 50 (12): 5919–5926.
APA
Zeitz, C., Labs, S., Lorenz, B., Forster, U., Uksti, J., Kroes, H. Y., De Baere, E., et al. (2009). Genotyping microarray for CSNB-associated genes. INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 50(12), 5919–5926.
Vancouver
1.
Zeitz C, Labs S, Lorenz B, Forster U, Uksti J, Kroes HY, et al. Genotyping microarray for CSNB-associated genes. INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE. 2009;50(12):5919–26.
MLA
Zeitz, Christina, Stephan Labs, Birgit Lorenz, et al. “Genotyping Microarray for CSNB-associated Genes.” INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE 50.12 (2009): 5919–5926. Print.
@article{939699,
  abstract     = {PURPOSE. Congenital stationary night blindness (CSNB) is a clinically and genetically heterogeneous retinal disease. Although electroretinographic (ERG) measurements can discriminate clinical subgroups, the identification of the underlying genetic defects has been complicated for CSNB because of genetic heterogeneity, the uncertainty about the mode of inheritance, and time-consuming and costly mutation scanning and direct sequencing approaches.
METHODS. To overcome these challenges and to generate a time-and cost-efficient mutation screening tool, the authors developed a CSNB genotyping microarray with arrayed primer extension (APEX) technology. To cover as many mutations as possible, a comprehensive literature search was performed, and DNA samples from a cohort of patients with CSNB were first sequenced directly in known CSNB genes. Subsequently, oligonucleotides were designed representing 126 sequence variations in RHO, CABP4, CACNA1F, CACNA2D4, GNAT1, GRM6, NYX, PDE6B, and SAG and spotted on the chip.
RESULTS. Direct sequencing of genes known to be associated with CSNB in the study cohort revealed 21 mutations (12 novel and 9 previously reported). The resultant microarray containing oligonucleotides, which allow to detect 126 known and novel mutations, was 100\% effective in determining the expected sequence changes in all known samples assessed. In addition, investigation of 34 patients with CSNB who were previously not genotyped revealed sequence variants in 18\%, of which 15\% are thought to be disease-causing mutations.
CONCLUSIONS. This relatively inexpensive first-pass genetic testing device for patients with a diagnosis of CSNB will improve molecular diagnostics and genetic counseling of patients and their families and gives the opportunity to analyze whether, for example, more progressive disorders such as cone or cone-rod dystrophies underlie the same gene defects. (Invest Ophthalmol Vis Sci. 2009;50:5919-5926) DOI:10.1167/iovs.09-3548},
  author       = {Zeitz, Christina and Labs, Stephan and Lorenz, Birgit and Forster, Ursula and Uksti, Janne and Kroes, Hester Y and De Baere, Elfride and Leroy, Bart and Cremers, Frans PM and Wittmer, Mariana and van Genderen, Maria M and Sahel, Jose-Alain and Audo, Iabelle and Poloschek, Charlotte M and Mohand-Said, Saddek and Fleischhauer, Johannes C and Huffmeier, Ulrike and Moskova-Doumanova, Veselina and Levin, Alex V and Hamel, Christian P and Leifert, Dorothee and Munier, Francis L and Schorderet, Daniel F and Zrenner, Eberhart and Friedburg, Christoph and Wissinger, Bernd and Kohl, Susanne and Berger, Wolfgang},
  issn         = {0146-0404},
  journal      = {INVESTIGATIVE OPHTHALMOLOGY \& VISUAL SCIENCE},
  keyword      = {ELECTRORETINOGRAM,RICH,COMPLETE FORM,FAMILIES,CACNA1F MUTATIONS,MUTATION DETECTION,STATIONARY NIGHT BLINDNESS,ARRAYED PRIMER EXTENSION,EXPRESSION,DYSTROPHY},
  language     = {eng},
  number       = {12},
  pages        = {5919--5926},
  title        = {Genotyping microarray for CSNB-associated genes},
  url          = {http://dx.doi.org/10.1167/iovs.09-3548},
  volume       = {50},
  year         = {2009},
}

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