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W. Li, R. Sui, G. Chen, F. Xu, Q. Zheng, P. Zhao, Y. Chen, X. Dai, F. Kong, J.-J. Pang; Screening of Gene Mutations in Chinese Patients With Leber Congenital Amaurosis by Arrayed Primer Extension. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1654.
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© ARVO (1962-2015); The Authors (2016-present)
Leber congenital amaurosis (LCA) is one of the most common forms of congenital blindness in childhood. To date, mutations in 15 genes were found to be associated with LCA. To accelerate mutant genes detection, arrayed primer extension (APEX), a new genotyping and resequencing technology that combines the efficiency of microarray format with the advantages of Sanger dideoxy sequencing has been employed. This pilot study is to test whether the known DNA changes reported in western LCA populations also occur in Chinese LCA patients.
DNA samples from 22 clinically diagnosed Chinese patients with LCA and 10 normal controls were analyzed using genotyping microarray based on the APEX technology (Zernant et al., 2005).
Four types of DNA changes that may cause LCA were detected. They were identified in 9 of 22 (40.9%) patients. The most common allele change in our LCA sample was CRB1 c.2306G>A (p.R769H), with a frequency of 4/44. Other detected allele changes included GUCY2D c.974T>C (p.L325P, 1/44), RPE65 c.11+5G>A (2/44), and CRB1 c.2714G>A (p.R905Q, 1/44). None of above DNA variations was found among 20 normal control alleles. Three SNPs, the AIPL1 c.268G>C p.D90H, GUCY2D c.154G>T p.A52S and RPGRIP1 c.574A>G p.K192E, were frequently detected in both LCA and normal control DNA samples.
Genotyping microarray appears to be an efficient and economic tool for preliminary screening of Chinese LCA patients and may benefit those with RPE65 mutations for future clinical trial in China. Our data suggest that the c.2306G>A p.R769H (CRB1) variation be the most common allele change, however our sample size is currently too small to draw any firm conclusion. More samples from both patients and normal controls are under examination to confirm if these allele changes are disease-related. Direct sequencing is also underway to verify whether there are any false positive findings.
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