title: Immunochromatographic assay for quantitative and sensitive detection of hepatitis B virus surface antigen using highly luminescent quantum dot-beads.
abstract: Hepatitis B virus infection is one of the major causes of hepatitis, liver cirrhosis and liver cancer. In this study, we used highly luminescent quantum dot-beads (QBs) as signal amplification probes in the sandwich immunochromatographic assay (ICA) for ultrasensitive and quantitative detection of hepatitis B virus surface antigen (HBsAg) in human serum. Various parameters that influenced the sensitivity and stability of the QB-based ICA (QB-ICA) sensor were investigated. Two linear independent regression equations for detection of serum HBsAg were expressed with Y=0.3361X-0.0059 (R(2)=0.9983) for low HBsAg concentrations between 75 pg mL(-1) and 4.8 ng mL(-1), and Y=0.8404 X-2.9364 (R(2)=0.9939) for high HBsAg concentrations in the range from 4.8 ng mL(-1) to 75 ng mL(-1). The detection limit of the proposed ICA sensor achieved was 75 pg mL(-1), which is much higher than that of the routinely-used gold nanoparticle based ICA. The intra- and inter-assays recovery rates for spiked serum samples at HBsAg concentrations of 75 pg mL(-1), 3.75 ng mL(-1) and 18.75 ng mL(-1) ranged from 90.14% to 97.6%, and coefficients of variation were all below 7%, indicating that the QB-ICA sensor has an acceptable accuracy for HBsAg detection. Additionally, the quantitative method developed showed no false positive results in an analysis of 49 real HBsAg-negative serum samples, and exhibited excellent agreement (R(2)=0.9209) with a commercial chemiluminescence immunoassay kit in identifying 47 HBsAg-positive serum samples. In summary, due to its high fluorescence intensity, the sandwich QB-ICA sensor is a very promising point-of-care test for rapid, simple and ultrasensitive detection of HBsAg, as well as other disease-related protein biomarkers.
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China.
authors.forename: Yaofeng
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: The Second Affiliated Hospital, Nanchang University, Nanchang 330000, PR China.
authors.forename: Fen
authors.initials: F
authors.lastname: Fu
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China. Electronic address: HengyiXu@ncu.edu.cn.
authors.forename: Hengyi
authors.initials: H
authors.lastname: Xu
authors.affiliation: The Second Affiliated Hospital, Nanchang University, Nanchang 330000, PR China.
authors.forename: Jiaofeng
authors.initials: J
authors.lastname: Lv
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, PR China. Electronic address: xiongyonghua@ncu.edu.cn.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Xiong
authors.affiliation: Ocean NanoTech, LLC., San Diego, CA 92126, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Wang
mesh_terms: D019187:Cadmium Compounds / Q000737:chemistry; D002846:Chromatography, Affinity; D006514:Hepatitis B Surface Antigens / Q000097:blood*; D006801:Humans; D049449:Luminescence; D045663:Quantum Dots / Q000737:chemistry*; D018036:Selenium Compounds / Q000737:chemistry; D013440:Sulfides / Q000737:chemistry; D017967:Zinc Compounds / Q000737:chemistry
chemical_list: D019187:Cadmium Compounds; D006514:Hepatitis B Surface Antigens; D018036:Selenium Compounds; D013440:Sulfides; D017967:Zinc Compounds; C058667:cadmium selenide; C031238:zinc sulfide
keywords: Hepatitis B virus surface antigen (HBsAg); Immunochromatographic assay; Quantitative determination; Quantum dot-beads