摘要
本研究旨在建立一种定量检测血清中CA19-9含量的时间分辨荧光免疫层析检测方法。采用双抗体夹心法与荧光免疫层析技术,以羧基荧光微球和NC膜为载体将CA19-9配对抗体进行标记和包被,制备CA19-9检测试纸条。通过标记、包被抗体量对工艺进行优化,并通过线性范围、最低检出限、精密性等性能指标对CA19-9时间分辨荧光层析检测方法进行评价。最终确定20μL荧光微球的标记抗体量为80μg,检测线包被抗体浓度为1.5 mg/mL时,检测时间为15 min,线性范围为12.5–800 U/mL,最低检出限为6.32 U/mL,批内精密性与批间精密性均小于15%,平均回收率为101%,与罗氏电化学发光检测试剂盒平行检测50份临床样本,两者相关系数为0.980 6。初步建立了定量检测血清中CA19-9的荧光免疫层析检测方法,有较好的临床应用前景。
To establish a time-resolved fluorescence immunochromatographic assay for quantitative determination of carbohydrate antigen 19-9(CA19-9) in serum, we prepared CA19-9 test strips by integrating double-antibody sandwich method and fluorescence immunochromatography technique. Carboxy fluorescent microspheres and nitrocellulose membrane were used as carriers for labeling and coating CA19-9 pairing antibodies. We optimized the process by adjusting the amount of labeling and coating antibody. According to the linear range, lowest detection limit and precision, We evaluated the time-resolved fluorescence immunochromatographic assay of CA19-9. When the amount of labeled antibody was 80 μg for 20 μL fluorescent microspheres, and the concentration of coated antibody on the test line was 1.5 mg/mL, the optimal reaction time was 15 minutes. Assay linear range was 12.5 to 800 U/mL and the minimum detection limit was 6.32 U/mL. The Within-run and between-run coefficient of variation were less than 15%. Average recovery rate was 101%. By detecting 50 clinical samples in parallel with Roche electrochemical luminescence detection kit, correlation coefficient was 0.980 6. The experiment, initially established a fluorescence immunochromatographic detection method to quantitative detection of serum CA19-9, which has a good clinical application prospect.
作者
王云龙
米亚双
李玉林
王继创
程蕾
闫生辉
邓黎黎
Yunlong Wang;Yashuang Mi;Yulin Li;Jichuang Wang;Lei Cheng;Shenghui Yan;and Lili Deng(College of Life Sciences, Henan Normal University, Xinxiang 453002, Henan, China;Henan Biotechnology Research Center, Zhengzhou 450121, Henan, China;Zhengzhou Technical College, Zhengzhou 450121, Henan, China;Biomarker Quantification Henan Engineering Laboratory, Zhengzhou 450121, Henan, China)
出处
《生物工程学报》
CAS
CSCD
北大核心
2018年第6期1012-1018,共7页
Chinese Journal of Biotechnology
基金
河南省科技攻关项目(No.162102310008)资助~~
关键词
CA19-9
荧光免疫层析
定量检测
快检
carbohydrate antigen 19-9
fluorescence immunochromatography
quantitative detection
rapid test