CAJ | 학술논문

目的采用时间分辨荧光免疫分析技术(TRFIA)建立高灵敏的恶性肿瘤相关糖类抗原(CA)242的快速全自动检测方法并进行初步应用.方法以抗CA242单克隆抗体601#包被96孔微孔板,以Eu3+-N2-[p-异氰酸苄基]-二乙烯三胺四乙酸标记抗CA242单克隆抗体242#,发光增强系统为以β-二酮体为主的增强液.采用平衡饱和法建立CA242 TRFIA,采用Auto DELLA1235系统考核CA242 TRFIA,并进行健康人和胃癌患者血清样本的检测.结果该方法的批内和批间CV分别为2.92%和5.36%,平均回收率为98.75%,灵敏度为0.2 kU/L,可测范围为0.2～200 kU/L,最高浓度点计数的20%、50%、80%对应的效应点值(ED20、ED50、ED80)分别为25.2 kU/L、53.3 kU/L和112.5 kU/L.CASO、CA19-9对CA242分别有15%和32%的交叉反应,癌胚抗原(CEA)、甲胎蛋白(AFP)、CA125对CA242 TRFIA均无交叉反应.Eu3+标记242#抗体于-20℃下保存6个月免疫反应性基本无损,同批试剂连续6个月应用分析结果稳定,检测健康体格检查人员的血清,样品的平均值为(6.52±5.15)kU/L,建议阳性阈值为17 kU/L,临床结果与酶联免疫吸附测定(ELISA)的结果相符,r为0.916.116例胃癌患者血清CA242的阳性率为26.7%.结论 CA242 TRFIA的灵敏度高、稳定性好,可用于血清CA242含量的免疫测定.
목적 채용시간분변형광면역분석기술(TRFIA)건립고령민적악성종류상관당류항원(CA)242적쾌속전자동검측방법 병진행초보응용.방법 이항CA242단극륭항체601#포피96공미공판,이Eu3+-N2-[p-이청산변기]-이을희삼알사을산표기항CA242단극륭항체242#,발광증강계통위이β-이동체위주적증강액.채용평형포화법건립CA242 TRFIA,채용Auto DELLA1235계통고핵CA242 TRFIA,병진행건강인화위암환자혈청양본적검측.결과 해방법 적비내화비간CV분별위2.92%화5.36%,평균회수솔위98.75%,령민도위0.2 kU/L,가측범위위0.2～200 kU/L,최고농도점계수적20%、50%、80%대응적효응점치(ED20、ED50、ED80)분별위25.2 kU/L、53.3 kU/L화112.5 kU/L.CASO、CA19-9대CA242분별유15%화32%적교차반응,암배항원(CEA)、갑태단백(AFP)、CA125대CA242 TRFIA균무교차반응.Eu3+표기242#항체우-20℃하보존6개월면역반응성기본무손,동비시제련속6개월응용분석결과 은정,검측건강체격검사인원적혈청,양품적평균치위(6.52±5.15)kU/L,건의양성역치위17 kU/L,림상결과 여매련면역흡부측정(ELISA)적결과 상부,r위0.916.116례위암환자혈청CA242적양성솔위26.7%.결론 CA242 TRFIA적령민도고、은정성호,가용우혈청CA242함량적면역측정.
Objective CA242 is valuable in the screening, diagnosis and prognosis prediction in pancreatic and colorectal tumors. The aim of this study was to establish a two-site time-resolved fluoroimmunoassay (TRFIA) of CA242 based on the sandwich technique and to study its clinical usefulness. Methods By the equilibrium method, the monoclonal antibodies (McAb) directed against a specific antigenic site on the CA242 was No. 601#, and the europium labelled McAb prepared from the europium chelate N-(p-iso-thiocyanatobenzyl)-diethylenetriamine-N,N,N,N-tetraacetic acid directed against a different antigenic site on the CA242 molecule was No.242#. The luminescent enhancement system contained mainly 2-naphthoyltrifluoroacetone. CA242 in sera from patients and healthy controls were determined by CA242-TRFIA by auto DELFIA1235 system. Results The intra-and inter-assay CV of the CA242-TRFIA were 2.92% and 5.36%, respectively, and the recovery rate was 98.75%. The sensitivity was 0.2 kU/L and the measurement range was 0.2-200 kU/L. The cross-reacting rates with CA50 and CA19-9 were 15% and 32%, respectively, while those with carcino-embryonic antigen (CEA), alpha fetoprotein (AFP) and CA125 were negligible. The europium labelled No.242# was at least stable for six months at-20℃. Serum samples from healthy blood donors were analyzed, resulting in a mean value of (6.52±5.15) kU/L, with an estimated reference limit ≤17 kU/L. The linear correlation of TRFIA and ELISA measurements resulted in a correlation coefficient of 0.916. The observed positive rate of CA242 in 116 gastric cancer cases was 26.7%. Conclusion The newly developed CA242-TRFIA technique was highly sensitive and stable in the immuno-determination of serum CA242.