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目的:分析胆汁泡相糖蛋白的特征并评估其ELISA药盒临床应用的可行性。方法:应用凝集素探针介导的点印迹法分析泡蛋白糖链的组成与结构,利用酶解方法测定泡蛋白各组分的构成,并分析其氨基酸组成。利用Westernblot法分析胆汁与血清泡蛋白的同源性,根据ELISAkit测定泡蛋白的含量分布,并以ROC曲线判别ELISAkit的临床应用价值。结果:化学分析泡蛋白由多肽和寡糖组成,其中多肽分子量约为21u,糖链约占总蛋白含量的37.3%,为核心岩藻糖丰富的多天线复杂型聚糖。泡蛋白具胆汁与血清分布两相性,胆固醇性结石组胆囊胆汁和血清33.5u泡蛋白含量分别为(213.4±70.1)μg/ml、(179.8±97.9)μg/ml,明显高于胆色素性结石症组和正常对照组(P<0.05),而后两者间差异无显著性(P>0.05)。ELISAkit判断胆固醇性结石的胆汁相敏感度和特异度分别为85%和90%,而血清相为61.7%和93.3%。结论:胆汁泡蛋白是一种新认识的胆结石促成核因子,其在不同人群中具有表达谱的异质性,ELISAkit辅助诊断胆石症具有一定的临床应用价值,但尚需更多病例的积累和验证。
Objective: To analyze the characteristics of bile bubble glycoprotein and evaluate the feasibility of its clinical application of ELISA kit. Methods: The composition and structure of the protein chains were analyzed by dot blotting mediated by lectin probe. The composition of each component of the protein was determined by enzymolysis method and its amino acid composition was analyzed. Western blot was used to analyze the homology of bile and serum protein, the content of vesicle was determined by ELISA kit, and the clinical value of ELISA kit was determined by ROC curve. Results: The chemical analysis of the proteolysin consisted of polypeptides and oligosaccharides, in which the molecular weight of the polypeptide was about 21u, the sugar chain accounted for about 37.3% of the total protein content, and was a core fucose-rich multi-antenna complex glycan. Bovine protein and bilirubin were distributed in two phases. The gallbladder bile and serum 33.5u protein in biliary calculi group were (213.4 ± 70.1) μg / ml and (179.8 ± 97.9) μg / ml, respectively, which were significantly higher than those in biliary pigment stones Disease group and normal control group (P <0.05), but there was no significant difference between the two groups (P> 0.05). The sensitivity and specificity of ELISA kit for the determination of cholesterol in gallstone were 85% and 90%, respectively, while the sera were 61.7% and 93.3% respectively. Conclusions: Biliary soaking protein is a newly recognized gallstone promoting factor. It has heterogeneity of expression profile in different populations. ELISA kit may be of clinical value in diagnosing cholelithiasis, but more cases need to be accumulated And verification.