摘要
目的建立可用于血清蔓陀罗凝集素(DSA)-γ-谷氨酰基转肽酶(GGT)检测的酶联免疫吸附法(ELISA),并探讨其在原发性肝癌(PHC)诊断中应用价值。方法制备抗DSA-GGT单克隆抗体(McAb),经蛋白G亲和层析柱色谱纯化后进行生物素标记,构建血清DSA-GGT生物素-链霉亲和素ELISA检测方法。用建立的方法对健康者、PHC患者和非PHC患者进行DSA-GGT检测,并进行方法学评价;采用受试者工作曲线(ROC曲线)确定DSA-GGT诊断PHC的临界值。结果所构建ELISA检测方法的最低检测限为2μg/L,平均批内、批间变异分别为8.9%和11.5%。健康者血清DSA-GGT水平呈正态分布,参考范围为(1.50±0.51)μg/L;经ROC曲线分析,确定其诊断PHC的临界值为3.25μg/L;其诊断PHC灵敏度为66.7%(28/40)、特异度为91.8%(112/122)。结论所建立的血清DSA-GGT生物素-链霉亲和素ELISA检测方法具有良好方法学性能,其诊断PHC的灵敏度、特异度良好,为PHC实验室诊断提供了新的方法。
Objective To establish an enzyme-linked immunosorbent assay （ELISA） for the detection of datura stramonim （DSA）-Y-glutamyltransferase（GGT） and investigate its clinical significance for the diagnosis of primary hepatic carcinoma（PHC）. Methods Anti-DSA-GGT monoclonal antibodies（McAb） was prepared and labeled with biotin,after being purified by protein G- sepharose affinity chromatography. And avidin-biotin ELISA for measurement of serum DSA-GGT was established and evaluated by detecting serum DSA-GGT in healthy subjects,patients with or without PHC. Cut-off value of DSA-GGT for the diagnosis of PHC was determined by receiver operating characteristic curve （ROC curve）. Results The minimum detection limit of serum DSA-GGT by avidin-biotin ELISA was 2 μg/L. Intra- and inter-assay coefficients of variation were 8.9 % and 11.5 %respectively. The distri- bution of DSA-GGT values showed Gaussian distribution and the reference range was（1.50±0.51）μg/L. Optimal cut-off value was 3.25 μg/L for the diagnosis of PHC,determined by ROC curve, with diagnostic sensitivity and specificity of 66.7% （28/40） and 91.8%（112/122） ,respectively. Conclusion The convenient avidin-biotin ELISA method was successfully established,with fine re- producibility and reliability. It might be a potential tool for the diagnosis of PHC to achieve higher sensitivity and specificity.
出处
《国际检验医学杂志》
CAS
2012年第22期2714-2716,2718,共4页
International Journal of Laboratory Medicine
