CAJ | 학술논문

目的建立测定乙酰肝素酶(HPA)的生物素-链亲和素系统(BSA) TRFIA并进行初步应用.方法采用部分基因重组的鼠抗人HPA单克隆抗体包被微孔板条,加入HPA标准品和生物素标记的HPA,两者竞争性结合HPA单克隆抗体,再利用铕标记链亲和素(Eu3+-SA)作为示踪物,建立竞争法BSA-TRFIA.利用该方法检测健康人血清样本(n=32)和肿瘤患者血清样本(n=54)HPA水平,并比较TRFIA结果与ELISA结果的相关性.数据分析采用两样本t检验或t’检验、直线相关分析.结果该法测定HPA的灵敏度为0.33 iug/L,批内和批间CV分别为5.29%和7.54%,平均回收率为105.5%,标准曲线范围为O-1 000 txg/L.健康人血清样本测定结果为(2.03±1.47)p.g/L,患者血清样本为(22.13_+7.38) tg/L(t’=19.388.P<O.01).该法测定值与ELISA结果相关(r=0.979,p<0.01).结论应用BSA-TRFIA建立的HPA测定法是一种灵敏度高,可测范围广,稳定,高效的定量检测方法.
목적 건립측정을선간소매(HPA)적생물소-련친화소계통(BSA) TRFIA병진행초보응용.방법 채용부분기인중조적서항인HPA단극륭항체포피미공판조,가입HPA표준품화생물소표기적HPA,량자경쟁성결합HPA단극륭항체,재이용유표기련친화소(Eu3+-SA)작위시종물,건립경쟁법BSA-TRFIA.이용해방법검측건강인혈청양본(n=32)화종류환자혈청양본(n=54)HPA수평,병비교TRFIA결과여ELISA결과적상관성.수거분석채용량양본t검험혹t’검험、직선상관분석.결과 해법측정HPA적령민도위0.33 iug/L,비내화비간CV분별위5.29%화7.54%,평균회수솔위105.5%,표준곡선범위위O-1 000 txg/L.건강인혈청양본측정결과위(2.03±1.47)p.g/L,환자혈청양본위(22.13_+7.38) tg/L(t’=19.388.P<O.01).해법측정치여ELISA결과상관(r=0.979,p<0.01).결론 응용BSA-TRFIA건립적HPA측정법시일충령민도고,가측범위엄,은정,고효적정량검측방법.
Objective To establish a novel TRFIA for the measurement of heparanase (HPA) in serum samples,and investigate its clinical application.Methods The micro-pore plate wells were first coated with partially recombinant murine anti-human HPA monoclonal antibody.Biotin-labeled recombinant HPA protein was then used to compete with HPA in serum samples,and the prepared europium (III)-labeled streptavidin (Eu3+-SA) was used as signal readout for establishing the BSA-TRFIA assay.Using this assay,the serum HPA levels in healthy subjects (n=32) and tumor patients (n=54) were measured.The results of BSA-TRFIA were compared with those of ELISA.Two-sample t test (or t' test),and linear correlation analysis were used to analyze the data.Results The sensitivity of BSA-TRFIA for measuring HPA was 0.33 ug/L.The CV values for intra-batch and inter-batch were 5.29% and 7.54%,respectively.The average recovery rate was 105.5%.The standard curve range was 0-1 000 ug/L.The serum HPA level measured by the BSA-TRFIA method in healthy subjects was (2.03_+ 1.47) Iug/L.In tumor patients,the HPA level was significantly higher:(22.13_+7.38) ug/L (t'=19.388,P<O.01).The HPA values measured by the BSATRFIA method were in good agreement with those by ELISA method (r=0.979,P<0.01).Conclusion BSA-TRFIA is a highly sensitive and stable method with efficient quantization and wide dynamic range for the measurement of HPA.