PCR-SSCP技术检测2型糖尿病患者脂联素基因3号外显子编码区突变

PCR-SSCP and DNA direct sequencing analysis in detecting mutation in exon 3 coding region of adiponectin gene

目的 观察人脂联素 (apM 1)基因 3号外显子编码区突变情况并探讨其与 2型糖尿病的关系。方法 选取 4 35例无亲缘关系的中国安徽地区汉族人 ,其中正常糖耐量(NGT)者 15 0例 ,2型糖尿病 (T2DM )者 2 85例。采用聚合酶链反应 单链构象多态性 (PCR SSCP)方法对apM1基因突变进行初筛分析后 ,对不同带型的标本进行DNA直接测序。结果 ①SSCP分析共发现 3种异常带型 ,测序后知为H14 2P、R2 2 1S及H2 4 1P3种杂合突变。②apM1基因H14 2P及R2 2 1S突变在T2DM组与NGT组间频率差异无统计学意义 (P >0 0 5 ) ,H 2 4 1P突变在T 2DM组中的频率明显高于NGT组 (P =0 0 38)。结论 apM1基因 3号外显子编码区存在 3种突变 ,其中H2 4 1P突变是 2型糖尿病的危险因素。

Objective To investigate the impact of the mutation in exon 3 coding region of adiponectin gene(apM1) on type 2 diabetes (T2DM) in Chinese Hans.Methods The study population consisted of 435 Chi-[FQ(13。46,X-WZ] nese Hans residents in Anhui province including 150 subjects with normal glucose tolerance (NGT) and 285 with T2DM. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to determine whether variation exist in apM1 gene.Then, exact mutation was performed by automated DNA direct sequencing.Results ①Three types of abnormal strands were found in SSCP analysis: H 142 P,R221S and H 241 P mutations in heterozygotes revealed by DNA direct sequencing.②The frequency of H142P and R221S mutations was not different between T2DM and NGT groups statistically (P>0.05).And T 2 DM group had more H 241 P mutation than NGT group(P=0.038).Conclusion There were three mutations in exon 3 coding region of apM1 gene, among which H241P mutation was associated with T2DM.