CAJ | 학술논문

目的：研究山东地区甲状腺发育不全（TD）的先天性甲状腺功能减退（CH）患儿甲状腺激素受体β（TRβ）基因突变类型及特点，为基因诊断及产前诊断提供理论依据。方法通过山东省新生儿筛查系统选取山东省60例 TD 的 CH 病例，提取患儿外周静脉血白细胞全基因组 DNA，用8对序列特异性引物对 TRβ基因的第6-12外显子进行 PCR 扩增，扩增后运用一代测序法（Sanger 测序法）对 PCR 产物直接进行测序。测序结果用 DNA MAN 软件与 TRβ基因的参考序列[美国国立生物技术信息中心（ NCBI）Reference Sequence：NC 000003.12]进行比对，结合比对结果分析是否存在基因突变。结果分析外显子测序结果，在60例患者中并未发现 TRβ基因突变，但在第9外显子和第6外显子区分别发现1个单核苷酸多态性（ SNP）位点（rs3752874，c.735C ﹥ T；rs79220627，c.162G ﹥ A），经分析，这2个 SNP 位点均为同义突变（ Phe→Phe；Ser→Ser），并未引起相应氨基酸发生改变。结论山东地区 TD 的 CH 患者中 TRβ基因突变率低，可能不是该地区TD 的 CH 患者的主要突变类型。
목적：연구산동지구갑상선발육불전（TD）적선천성갑상선공능감퇴（CH）환인갑상선격소수체β（TRβ）기인돌변류형급특점，위기인진단급산전진단제공이론의거。방법통과산동성신생인사사계통선취산동성60례 TD 적 CH 병례，제취환인외주정맥혈백세포전기인조 DNA，용8대서렬특이성인물대 TRβ기인적제6-12외현자진행 PCR 확증，확증후운용일대측서법（Sanger 측서법）대 PCR 산물직접진행측서。측서결과용 DNA MAN 연건여 TRβ기인적삼고서렬[미국국립생물기술신식중심（ NCBI）Reference Sequence：NC 000003.12]진행비대，결합비대결과분석시부존재기인돌변。결과분석외현자측서결과，재60례환자중병미발현 TRβ기인돌변，단재제9외현자화제6외현자구분별발현1개단핵감산다태성（ SNP）위점（rs3752874，c.735C ﹥ T；rs79220627，c.162G ﹥ A），경분석，저2개 SNP 위점균위동의돌변（ Phe→Phe；Ser→Ser），병미인기상응안기산발생개변。결론산동지구 TD 적 CH 환자중 TRβ기인돌변솔저，가능불시해지구TD 적 CH 환자적주요돌변류형。
Objective To study the thyroid hormone receptor β(TRβ)gene mutation types and characteristics in children with congenital hypothyroidism(CH)and thyroid dysgenesis(TD)from Shandong Province,and to provide theoretical basis for gene diagnosis and prenatal diagnosis. Methods Sixty cases of TD patients of which genomic DNA were isolated from peripheral blood leukocytes were selected by neonatal screening system in Shandong Province. The exon 6 to 12 of TRβ gene were amplified with 8 pairs of sequence specific primers using PCR and the first generation of sequencing method(Sanger method)to detect mutation. The sequencing results were compared with the TRβ gene reference sequence[National Center for Biotechnology Information(NCBI)Reference Sequence:NC 000003. 12]to see whether there was a mutation. Results Analysis of TRβ in 60 cases of CH patients with TD revealed no mutation was demonstrated in exons 6 - 12,but 2 single nucleotide polymorphism(SNP)( rs 3752874,c. 735C ﹥ T;rs79220627, c. 162G ﹥ A)were detected. Through the analysis,the 2 SNP were all synonymous mutations(Phe→Phe;Ser→Ser), without the change of the amino acids. Conclusions TRβ mutation rate is very low,which may not be the main mutation type in CH patients with TD in Shandong Province.