转铁蛋白基因多态性对毛细管电泳法检测血清糖缺失性转铁蛋白的影响

The Influence of Transferrin Gene Polymorphism on Measuring Carbohydrate Deficient Transferrin by Capillary Electrophoresis

目的研究转铁蛋白(Tf)基因多态性对毛细管电泳检测血清糖缺失性转铁蛋白(CDT)的影响,探讨其临床应用价值。方法选择51例健康体检者,17例非酒精性肝病者,65例酗酒者作为研究对象,均为汉族成年男性。首先采用毛细管电泳(CE)获取样本的CDT电泳图,选取会影响毛细管电泳对血清中CDT含量检测并且在中国人群中较为常见的Tf基因变异体:Tf-Dchi(A>G)、Tf-D1(A>G)与Tf-B2(G>A),应用高分辨熔解曲线(HRM)初步检测所有标本的基因型,对HRM筛选出的样本进行测序验证。结果 133例标本经毛细管电泳后显示:127例峰分离良好,6例未能完全分离(健康组2例,非酒精性肝病3例,酗酒组1例),导致所得CDT结果不可信。133例样本经HRM分析,可疑突变样本经测序确认后,分别检出3例Tf-Dchi(A>G)杂合突变型,0例TfD1(A>G)突变型,0例Tf-B2(G>A)突变型,其余3例均为C型。Tf-Dchi、Tf-D1、Tf-B2突变型在被检测人群中出现概率分别为2.3%(3/133),0,0。结论 Tf基因多态性中Tf-Dchi型变异体会导致毛细管电泳中Tf异构体分离不佳,影响检测准确性。

Objective To determine the influence of isoforms of transferrin （Tf） on the detection of serum carbohydrate-deficient transferrin （CDT） by capillary electrophoresis （CE）. Methods A total of 51, 17, and 65 serum samples were collected from the healthy participants, the patients with non-alcoholic liver diseases, and the patients with alcoholic liver diseases, respectively. Serum CDT was measured by CE. The genotype of Tf of the samples without a good separation was further analyzed using high resolution melting （HRM） methods. Those with suspected mutation were confirmed by sequencing. Results Six samples showed incomplete separation （2 from the healthy participants, 3 from the patients with non-alcoholic liver diseases, 1 from the patient with alcoholic liver diseases）. Of the 133 serum samples, 2.3% （3/133） were detected with Tf-Dchi heterozygote. But no Tf-D1 and Tf-B2 heterozygote were identified. Conclusion Tf-Dchi variant influences the measurement of carbohydrate deficient Tf by CE, leading to unreliable results.