Establishment of time-resolved fluorescence immunochromatographic assay for detection of carbohydrate antigen 19-9.
- Author:
Yunlong WANG
1
;
Yashuang MI
1
;
Yulin LI
2
;
Jichuang WANG
2
;
Lei CHENG
2
;
Shenghui YAN
3
;
Lili DENG
4
Author Information
- Publication Type:Journal Article
- Keywords: carbohydrate antigen 19-9; fluorescence immunochromatography; quantitative detection; rapid test
- From: Chinese Journal of Biotechnology 2018;34(6):1012-1018
- CountryChina
- Language:Chinese
- Abstract: 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.
