Mutational screening of 10 genes in Chinese patients with microphthalmia and/or coloboma
Mutational screening of 10 genes in Chinese patients with microphthalmia and/or coloboma
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DOI：
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发表时间：
2009-12
期刊：
影响因子：
2.2
通讯作者：
Xiaohui Zhang;Shi-qiang Li;Xueshan Xiao;Xiao-yun Jia;Panfeng Wang;Huangxuan Shen;Xiangming Guo;Qingjiong Zhang
中科院分区：
文献类型：
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作者：
Xiaohui Zhang;Shi-qiang Li;Xueshan Xiao;Xiao-yun Jia;Panfeng Wang;Huangxuan Shen;Xiangming Guo;Qingjiong Zhang
目的对32例小眼球和/或眼球缺损患者的10个基因进行突变筛查。方法对32例无血缘关系的小眼症（9例先证者）和葡萄膜缺损（23例先证者）患者的基因组DNA进行制备。采用循环测序法检测10个基因的序列变异，包括BMP 4、VSX 2、BMP 4、GDF 6、OTX 2、RAX、SIX 3、SIX 6、SOX 2和LRP 6。采用限制性片段长度多态性（RFLP）或杂合酶-单链构象多态性（HA-SSCP）分析方法，对96名无关对照进行了进一步的分析。结果在10个基因中，在1例双侧小眼球合并单侧白内障患者的BMP 4基因中检测到一个新的c.751C>T（p.H251Y）基因。c.751C>T变异也存在于他健康的兄弟（可能是正常的父母之一）中。此外，在SIX 6中鉴定了一种新的c.608G>A（p.R203Q），作为优化实验条件的内部对照。内部对照来自一名患有典型无虹膜和PAX 6中鉴定的c.718C>T（p.R240X）突变的女孩，表明SIX 6中的c.608G>A变异不太可能在她的眼部表型中发挥作用。96例正常对照组中BMP 4的c.751C>T和SIX 6的c.608G>A均不存在。此外，还检测到16个核苷酸替换，包括8个已知的SNP和8个新的同义变化。结论虽然小眼畸形和/或缺损的遗传病因学尚不清楚，但相关基因的罕见变异，如SIX 6中的c.608 G>A和BMP 4中的c.751 C>T，可能不是病因。这些结果进一步强调了仔细的临床和遗传分析在突变-疾病关联中的重要性。
Purpose To screen ten genes for mutations in 32 Chinese patients with microphthalmia and/or coloboma. Methods Genomic DNA was prepared from 32 unrelated patients with microphthalmia (nine probands) and uveal coloboma (23 probands). Cycle sequencing was used to detect sequence variations in ten genes, including BMP4, VSX2, CRYBA4, GDF6, OTX2, RAX, SIX3, SIX6, SOX2, and LRP6. Variations were further evaluated in 96 unrelated controls by using restriction fragment length polymorphism (RFLP) or heteroduplex-single strand conformation polymorphism (HA-SSCP) analysis. Results In the ten genes, a novel c.751C>T (p.H251Y) in BMP4 was detected in a patient with bilateral microphthalmia and unilateral cataract. The c.751C>T variation is also present in his healthy brother (and possibly one of the normal parents). In addition, a novel c.608G>A (p.R203Q) in SIX6 was identified in an internal control for optimizing experimental conditions. The internal control was from a girl with typical aniridia and an identified c.718C>T (p.R240X) mutation in PAX6, suggesting the c.608G>A variation in SIX6 was unlikely to play a role in her ocular phenotype. The c.751C>T in BMP4 and the c.608G>A in SIX6 were not present in the 96 normal controls. In addition, 16 nucleotide substitutions, including eight known SNPs and eight new synonymous changes, were detected. Conclusions Although the genetic etiology for microphthalmia and/or coloboma is still elusive, rare variations in the related genes, such as c.608 G>A in SIX6 and c.751C>T in BMP4, may not be causative. These results further emphasize the importance of careful clinical and genetic analysis in making mutation-disease associations.