A fluorescence immunochromatography method for detection of human papillomavirus type 16 E6 and L1 proteins.
- Author:
Xin'er LIU
1
;
Yinzhen ZHAO
2
;
Nannan NIU
2
;
Lingke LI
2
;
Xueli DU
2
;
Jinxiang GUO
2
;
Yingfu ZHANG
3
;
Jichuang WANG
4
;
Yiqing ZHANG
5
;
Yunlong WANG
1
Author Information
- Publication Type:Journal Article
- Keywords: cervical cancer; human papillomavirus; rapid detection; time-resolved fluorescence immunochromatography assay (TRFIA)
- MeSH: Oncogene Proteins, Viral/immunology*; Humans; Chromatography, Affinity/methods*; Female; Human papillomavirus 16; Repressor Proteins/immunology*; Capsid Proteins/immunology*; Papillomavirus Infections/diagnosis*; Fluorescence; Uterine Cervical Neoplasms/virology*
- From: Chinese Journal of Biotechnology 2024;40(11):4266-4276
- CountryChina
- Language:Chinese
- Abstract: This study aims to establish a time-resolved fluorescence immunochromatography method for simultaneous determination of human papillomavirus (HPV) type 16 E6 and L1 protein concentrations. The amount of lanthanide microsphere-labeled antibodies, the concentration of coated antibodies, and the reaction time were optimized, and then a test strip for the simultaneous determination of the protein concentrations was prepared. The performance of the detection method was evaluated based on the concordance of the results from clinical practice. The optimal conditions were 8 μg and 10 μg of HPV16 L1 and E6-labeled antibodies, respectively, 1.5 mg/mL coated antibodies, and reaction for 10 min. The detection with the established method for L1 and E6 proteins showed the linear ranges of 5-320 ng/mL and 2-64 ng/mL and the lowest limits of detection of 1.78 ng/mL and 1.09 ng/mL, respectively. There was no cross reaction with human immunodeficiency virus (HIV), treponema pallidum (TP), or HPV18 E6 and L1 proteins. The average recovery rate of the established method was between 97% and 107%. The test strip prepared in this study showed the sensitivity, specificity, and diagnostic accuracy of 97.46%, 90.57%, and 95.32%, respectively, in distinguishing patients with cervical cancer and precancerous lesions from healthy subjects, with the area under the curve (AUC) of 0.980 1 and 95% Confidence Interval (CI) of 0.956 5 to 1.000 0. The time-resolved fluorescence immunochromatography combined with the test strips prepared in this study showed high sensitivity, high accuracy, simple operation, and rapid reaction in the quantitation of HPV16 E6 and L1 proteins. It thus can be used as an auxiliary method for the diagnosis and early screening of cervical cancer and precancerous lesions and the assessment of disease course.
