Rare Earth Fluorescent Probes for Rapid Detection of Hepatitis B Surface Antigen

doi:10.19894/j.issn.1000-0518.240055

Chinese Journal of Applied Chemistry ›› 2024, Vol. 41 ›› Issue (12): 1760-1769.DOI: 10.19894/j.issn.1000-0518.240055

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Rare Earth Fluorescent Probes for Rapid Detection of Hepatitis B Surface Antigen

Rui-Qi ZHANG¹, Yu CHANG², Zuo-Jia LIU², Xue-Song LI¹(), Li-Hua PAN²()

^1.Ministry of Education Key Laboratory of Advanced Structural Materials，College of Materials Science and Engineering，Changchun University of Technology，Changchun 130012，China
^2.Changchun Institute of Applied Chemistry，Chinese Academy of Sciences，Changchun 130022，China

Received:2024-02-22 Accepted:2024-10-16 Published:2024-12-01 Online:2025-01-02
Contact: Xue-Song LI,Li-Hua PAN
About author:lihuap@ciac.ac.cn；
lixuesong@ccut.edu.cn
Supported by:
Xinjiang Uygur Autonomous Region Science and Technology Department Key Research and Development Project(2021B03003);Jilin Provincial Science and Technology Department Key Research and Development Project(20224094094XY);Jilin Provincial Natural Science Foundation Upper-level Project(20230101330JC)

Abstract

Abstract:

In this study， a self-synthesized rare-earth chelator 4，4′-Bis（1″，1″，1″，2″，2″，3″，3″-7-fluoro-4″，6″-hexanedione-6″-yl）chlorosulfonated terphenyl （BHHCT） was used to prepare hepatitis B surface antibody markers， and a time-resolved fluorescence immunoassay for the rapid detection of hepatitis B surface antigen （HBsAg） was developed by applying lateral-flow immunochromatographic test strips and an accompanying time-resolved fluorescence detection system. The linear range was 0.2~500 ng/mL， the detection limit was 0.16 ng/mL， and the linear regression coefficient of determination （R²） was 0.9996. The intra-batch coefficient of variation was less than 5%， the inter-batch coefficient of variation was less than 8%， and the recoveries of the samples ranged from 94.3% to 107.8%. The method has the advantages of precision， rapidity， specificity and no need for specialized personnel to operate， which can meet the requirement of immediate detection of hepatitis B surface antigen.

Key words: Time-resolved fluorescence immunoassay, Lateral flow immunochromatography strip, Hepatitis B surface antigen, Point of care testing

CLC Number:

O655

Rui-Qi ZHANG, Yu CHANG, Zuo-Jia LIU, Xue-Song LI, Li-Hua PAN. Rare Earth Fluorescent Probes for Rapid Detection of Hepatitis B Surface Antigen[J]. Chinese Journal of Applied Chemistry, 2024, 41(12): 1760-1769.

Figures/Tables 11

Fig.1 （A）Schematic diagram of the composition and working principle of immunochromatographic test strips；（B） The experimental operation process and the time required for each step

Fig.2 （A） UV absorption spectra of BHHCT-Eu3+ and （B） emission spectra of Ab-BHHCT-Eu3+ at different mass concentration

Fig.3 UV-Vis spectra of antibodies dAb， BHHCT-Eu3+ and dAb-BHHCT-Eu3+

Fig.4 （A） Optimization of labeling reaction buffers with different pH；（B） Marking ratios for buffers with different pH；（C） Optimization of labeling reaction times；（D） Marking ratios for different reaction times

Fig.5 （A） Fluorescent test strips images for detection HBsAg solution samples with different drip volumes and （B） corresponding FT/FC values

Fig.6 （A） Fluorescent test strips images of TRFILF for detection of different mass concentrations of HBsAg and （B） FT/FC values；（C） Linear fitting curves of FT/FC values to HBsAg to mass concentration of HBsAg

Fig.7 （A） Specificity test results and （B） corresponding FT/FC values

Fig.8 Stability test result at （A） 4 and （B） 37 ℃

Table 1 Precision measurement results

ρ（HBsAg）/（ng·mL^-1）	Intra-assay CV/%（n=10）	Inter-assay CV/%（n=3）
2.0	4.6	7.7
20.0	2.4	5.3
250.0	2.6	5.1

Table 2 Accuracy measurement results

Added/（ng·mL^-1）	Found/（ng·mL^-1）	Recovery/%	CV/%（n=10）
2	2.16	108.0	3.9
20	18.86	94.3	2.2
250	245.62	98.2	2.5

Table 3 Comparison of the performance of this work with other methods for the detection of HBsAg

Method	Limit of detection/（ng·mL^-1）	Detection range/（ng·mL^-1）	Ref.
AuNP	2	5~200	［22］
ELISA	1	0~40	［26］
CdS nanowires	0.5	0.02~100	［26］
Modified europium nanoparticles	0.25	0.01~100	［34］
Ultramarine blue particles	0.37	1~50	［35］
Rare earth fluorescent probes	0.16	0.2~500	This work

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Rare Earth Fluorescent Probes for Rapid Detection of Hepatitis B Surface Antigen

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