基于微流控技术与酶促化学发光免疫技术检测血清CEA方法的建立及初步应用评价  

Establishment and preliminary evaluation of serum CEA detection based on microfluidic technology and chemiluminescence enzyme immunoassay

作　　者：梁臻龙[1] 王策[1] 李炯[2] 刘萍[3] 陈运霞[3] 王成彬[1] 

机构地区：[1]温州医科大学检验医学院生命科学学院,浙江温州325035 [2]中国科学院苏州纳米技术与纳米仿生研究所,江苏苏州215123 [3]中国人民解放军总医院临床检验科,北京100853

出　　处：《临床检验杂志》2015年第12期892-894,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的基于微流控技术和酶促化学发光免疫技术的原理建立一种用于检测癌胚抗原（CEA）的方法并进行初步应用评价。方法采用碳二亚胺（EDC）/N-羟基琥珀酰亚胺（NHS）化学偶联的方法将鼠抗人CEA包被于直径为1 mm的二氧化硅微球表面,将其装载于毛细玻璃管中,在微量注射泵的作用下,用双抗体夹心酶免疫法检测CEA,并进行方法学与临床应用评价。结果该方法可在30 min内完成CEA检测,最低检测浓度达0.54 ng/m L,与高浓度甲胎蛋白、间皮素无明显交叉反应,批内、批间CV为10.53%-14.22%,回收率为91.20%-104.65%,与电化学发光酶免疫分析结果差异无统计学意义（t=0.533,P=0.597）,且成正相关（r=0.980,P＆lt;0.01）。结论基于微流控和酶促化学发光免疫技术的原理在毛细玻璃管中检测CEA,操作简便、快速、灵敏度高且成本低,方法学性能良好。Objective To establish a method for the detection of serum carcinoembryonic antigen（CEA） based on microfluidic technology and chemiluminescence enzyme immunoassay, and evaluate its methodology and clinical application. Methods The mouse anti-human CEA antibody was coated on the surface of silica microspheres with a diameter of 1 mm by the coupling method with 1-（ 3- dimethylaminopropyl） -3-ethylcarbodiimide hydrochloride （EDC） and N-hydroxysuccinimide （NHS）. Then, the chemically modified mi- crospheres were loaded into the capillary tube, and CEA was detected based on the infusion by a syringe pump and a sandwich assay. Last, the established method was evaluated. Results Serum CEA could be detected by the established method within 30 min, and the lowest detection concentration of CEA was 0. 54 ng/mL. There was no obvious cross reaction between CEA and high concentration of alpha-fetoprotein, mesothelin and other tumor markers. The intra- and inter-assay coefficients of variation（CV） ranged from 10.53% to 14.22%, and the recovery rates ranged from 91.20% to 104.65%. There was no significant difference in the serum CEA levels detected by the established method and the electrochemiluminescence method（ t = 0.533, P = 0.597）, and there was positive correlation between them（ r = 0. 980, P 〈 0.01）. Conclusion The established method for the detection of serum CEA based on the microfluidics and chemiluminescence enzyme immunoassay is simple, rapid, highly sensitive and low cost, and has a good performance of methodology.

关 键 词：化学发光 癌胚抗原 方法学性能评价 

分 类 号：R446[医药卫生—诊断学]