DSA亲和色谱检测肝癌特异性γ-谷氨酰转移酶的临床应用

Clinical application of HS-GGT detected by a simple affinity chromatography method

目的应用建立的凝集素亲和色谱技术,分离血清肝癌特异性γ-谷氨酰转移酶(HS-GGT),并观察HS-GGT在正常人、良性肝病及原发性肝癌(PHC)患者中的变化,以期建立1种简便易行的PHC实验室诊断方法。方法PHC 64例、肝外肿瘤22例、良性肝病68例及正常对照血清21例,以神经胺酸酶(NMD)水解后直接用欧蔓陀罗凝集素(DSA)亲和色谱柱分离,首先用柱平衡液洗脱不结合组分,再用0.05 mol/L醋酸洗弱结合组分,最后用0.1 mol/L醋酸洗脱强结合组分,用连续监测法测定洗脱液的GGT活性,计算各组分所占比例,统计分析其临床应用价值。结果DSA强结合的GGTⅢ为PHC患者所特有,以其在总GGT所占比例大于2%为PHC诊断的Cutoff值,其诊断特异性为95.5%,灵敏度为85.8%,准确度为92.0%。结论该方法能较好地鉴别诊断PHC,较以往的其他检测方法更为简便,并有较高诊断灵敏度和特异性。

Purpose A simple method for detection of hepatoma-speeific gamma- glutamyltranspeptidase [HS-GGT] was set up by DSA affinity chromatography, and its clinical application was verified in diagnosis of primary hepatocellular careinoma[PHC] .Methods Serums were collected from 64 patients with PHC, 22 with extrahepatic tumor, 68 with benign liver diseases and 21 normal controls, and then analyzed by DSA-sepharose affinity chromatography after digestion by NMD. Three fractions were eluted and collected by equilibration buffer, 0.05 mol/L acetate and 0.1 mol/L acetate respectively. GGT activity in each fraction was tested by continuous monitoring method, and the diagnostic value of GGT in each fraction was assessed and compared statistically. Results DSA strongly bound GGT Ⅲ is specific for PHC, if we set the proportion of GGT Ⅲ 〉 2% as the cutoff value, the specificity, sensitivity and accuracy of GGT Ⅲ in diagnosis of PHC is 95.5%, 85.8% and 92.0% respectively. Conclusion The method is helpful for differential diagnosis of PHC, and more sensitive and specific with much simpler procedure compared with other methods.