He Wenbin, Lin Ge, Liang Ping, Cheng Dehua, Hu Xiao, Zhou Lihua, Xiong Bo, Tan Yueqiu, Lu Guangxiu, Li Wen
Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, Hunan 410078, China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2017 Aug 10;34(4):476-480. doi: 10.3760/cma.j.issn.1003-9406.2017.04.002.
To explore the genetic etiology of three families affected with split-hand/split-foot malformation (SHFM).
Peripheral venous blood samples from 21 members of pedigree 1, 2 members of pedigree 2, and 2 members of pedigree 3 were collected. PCR-Sanger sequencing, microarray chip, fluorescence in situ hybridization (FISH), real-time PCR, and next-generation sequencing were employed to screen the mutations in the 3 families. The effect of the identified mutations on the finger (toe) abnormality were also explored.
Microarray and real-time PCR analysis has identified a duplication in all patients from pedigrees 1 and 3, which have spanned FKSG40, TLX1, LBX1, BTRC, POLL and FBXW4 (exons 6-9) and LBX1, BTRC, POLL and FBXW4 (exons 6-9) genes, respectively. A missense mutation of the TP63 gene, namely c.692A>G (p.Tyr231Cys), was found in two patients from pedigree 2. FISH analysis of chromosome 10 showed that the rearrangement could fita tandem duplication model. However, next-generation sequencing did not identify the breakpoint.
The genetic etiology for three families affected with SHFM have been identified, which has provideda basis for genetic counseling and guidance for reproduction.
探究三个患有裂手/裂足畸形(SHFM)家庭的遗传病因。
采集了家系1的21名成员、家系2的2名成员和家系3的2名成员的外周静脉血样本。采用聚合酶链反应-桑格测序、微阵列芯片、荧光原位杂交(FISH)、实时聚合酶链反应和下一代测序技术对这3个家庭进行突变筛查。还探究了所鉴定突变对手指(脚趾)异常的影响。
微阵列和实时聚合酶链反应分析在来自家系1和家系3的所有患者中分别鉴定出一个重复,其分别跨越了FKSG40、TLX1、LBX1、BTRC、POLL和FBXW4(外显子6 - 9)基因以及LBX1、BTRC、POLL和FBXW4(外显子6 - 9)基因。在家系2的两名患者中发现了TP63基因的一个错义突变,即c.692A>G(p.Tyr231Cys)。对10号染色体的FISH分析表明该重排符合串联重复模型。然而,下一代测序未鉴定出断点。
已确定三个患有SHFM家庭的遗传病因,这为遗传咨询和生育指导提供了依据。