Identification of the susceptibility genes for nonsyndromic cleft lip and /or palate

1)九州大学口腔科学研究生院和鹿儿岛大学医学与口腔科学研究生院的伦理委员会批准了非综合征型唇腭裂（CLP）易感基因的鉴定。合作鉴定CLP易感基因也获得了印尼Harapan Kita Children and妇产医院伦理委员会的批准。2)在知情同意后，采用伦理委员会批准的方案，对113组患者和家长的血液样本进行了校正。在选择非综合征性CLP患者的多家系后，对6个多家系的指甲样本进行校正。从血样中提取DNA。采用直接测序法和焦磷酸测序法对GABAA受体基因和Fox2基因各标记进行基因分型。使用计算程序继续评估每个标记物与非综合征性CLP之间的关联。4)对Ellis-van Creveld综合征家族性病例，采用直接测序法对MSX1、EVC、EVC2基因各标记进行基因分型。所有22个外显子中的6个外显子未检测到EVC基因突变，后续研究将继续进行。5)对443例唇腭裂合并唇腭裂患者的复杂畸形进行分析。复杂性畸形发生率最高的是面腭裂（70%），其次是黏膜下腭裂（54.3%）和腭裂（47.4%）。其发生率高于CLA（15.3%）和CLP（18.4%）。

1) Approval o identification of the susceptibility genes for nonsyndromic cleft lip and/or palate (CLP) was obtained in the ethics committees in Graduate School of Dental Science in Kyushu University and Graduate School of Medical and dental Sciences in Kagoshima University. Also approval of collaboration for identification of the susceptibility genes for CLP was obtained in the ethics committee in Harapan Kita Children and Maternity Hospital in Indonesia.2) Correcting the blood samples from patient and parents was made in 113 trios after informed consent using protocol approved by the ethic committee. After selection of the multiples families with nonsyndromic CLP patients, the nail samples were corrected from 6 multiples families.3) DNA extraction was made from the blood samples. Genotyping of each marker of the GABAA receptor gene and Fox2 gene was carried out by the direct sequencing and the pyro-sequencing method. Evaluation the associations between each marker and nonsyndromic CLP was continued by using the computational program. Furthermore, selecting the genomic markers on the susceptibility chromosomal regions supposed from latest was made on the samples extracted from multiples families in Harapan Kita Hospital.4) On the familial cases of Ellis-van Creveld syndrome, genotyping of each marker of MSX1, EVC, and EVC2 genes was carried out by the direct sequencing method. No mutation of EVC gene has been detected in the 6 Exons of all 22 Exons, and further studies will be continued.5) Complicated malformation of cleft lip and palate was analyzed on 443 patients with CLP and facial cleft. Incidence complicated malformation was highest in facial cleft (70%), followed by submucous cleft palate (54.3%) and cleft palate (47.4%). They were found to be more frequent than CLA (15.3%) and CLP (18.4%).