血清中TIGAR的ELISA建立及初步临床试验

目的建立肿瘤蛋白53（TP53）诱导的糖酵解和凋亡的调控子（TIGAR）的定量酶联免疫吸附试验（ELISA）,并探讨血清中TIGAR定量测定在肿瘤患者诊断中的临床应用。方法构建TIGAR原核表达载体并诱导蛋白表达,纯化蛋白后免疫家兔获得多克隆抗体,以此兔抗TIGAR多克隆抗体为包被抗体,以生物素-辣根过氧化物酶标记链霉亲和素（SA-HRP）检测系统,建立TIGAR定量检测的双抗体夹心ELISA。测定40例肿瘤患者及40名对照者的血清TIGAR含量,并探讨其与肿瘤的关系。结果建立的TIGAR定量ELISA拟合曲线的相关系数（r）=0.99,肿瘤患者血清TIGAR含量高于对照组,差异有统计学意义（S=954.5,P〈0.01）。结论本研究成功建立了TIGAR定量ELISA,血清TIGAR含量测定对某些肿瘤的研究和临床检测可能有一定价值。

Establishment of an ELISA for measuring TIGAR and its preliminary clinical trial

Objective To establish an enzyme-linked immunosorbent assay（ELISA） for measuring tumor protein 53（TP53）-induced glycolysis and apoptosis regulator（TIGAR） and explore the clinical application of the measurement of TIGAR in the diagnosis of tumor diseases.Methods A recombinant plasmid expressing TIGAR protein was constructed,and the recombinant protein His-TIGAR was purified.The polyclonal antibody against TIGAR was prepared.With rabbit polyclonal anti TIGAR antiserum as coating antibody and biotin-streptavidin-horseradish peroxidase（SA-HRP） as detecting system,a double antibody sandwich ELISA was established for measuring TIGAR.The serum levels of TIGAR from 40 patients with tumor and 40 healthy subjects were measured and analyzed.Results The correlation coefficient（r） was 0.99 for TIGAR assay.The levels of TIGAR in patients with tumor were higher than those in healthy subjects,and it had statistical differences（S=954.5,P0.01）.Conclusions An ELISA for measuring the TIGAR in serum is successfully established.The determination of TIGAR in serum may be valuable for the research and diagnostic examination for certain tumors.